Methods of treating neurological, metabolic, and other disorders using enantiopure deuterium-enriched pioglitazone

ABSTRACT

The invention provides enantiopure deuterium-enriched pioglitazone, pharmaceutical compositions, and methods of treating neurological disorders, cancer, respiratory disorders, metabolic disorders, and other disorders using enantiopure deuterium-enriched pioglitazone. A preferred aspect of the invention provides methods of treating Alzheimer&#39;s disease, non-small cell lung cancer, hepatocellular carcinoma, and chronic obstructive pulmonary disease using enantiopure deuterium-enriched pioglitazone.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/211,488, filed Dec. 6, 2018 which is a divisional of U.S. patentapplication Ser. No. 15/109,533, filed Oct. 7, 2016 which is thenational stage of International (PCT) Patent Application serial numberPCT/US2015/011493, filed Jan. 15, 2015 which claims the benefit of andpriority to U.S. Provisional Patent Application Ser. No 61/927,708,filed Jan. 15, 2014, and to U.S. Provisional Patent Application Ser. No.61/931,808, filed Jan. 27, 2014; the contents of each of which arehereby incorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention provides enantiopure deuterium-enriched pioglitazone,pharmaceutical compositions, and methods of treating neurologicaldisorders, cancer, respiratory disorders, metabolic disorders, and otherdisorders using enantiopure deuterium-enriched pioglitazone.

BACKGROUND

Peroxisome proliferator-activated receptors (PPARs) are a group ofnuclear receptor proteins that function as transcription factorsregulating the expression of genes. There are three subtypes of thesereceptors, PPAR alpha, beta, and gamma. PPARs mainly regulate theexpression of genes involved in the regulation of lipid and carbohydratemetabolism. These receptors are also involved in the regulation ofinflammatory processes, reproduction, carcinogenesis, and otherphysiological processes in the body. Treatment of a variety of medicaldisorders (e.g., Alzheimer's disease, cancer, and chronic obstructivepulmonary disease) has been linked to modulating the activity (e.g.,activation) of certain PPARs.

Therapeutics that modulate PPARs have been commercialized for treatingmedical disorders, such as metabolic disorders. One such example ispioglitazone hydrochloride, which has been approved by the United StatesFood and Drug Administration as an adjunct to diet and exercise toimprove glycemic control in adults with type 2 diabetes mellitus inmultiple clinical settings. Pioglitazone hydrochloride is marketed underthe registered trademark ACTOS® and the prescribing information forACTOS® explains that pioglitazone is an agonist of PPAR gamma Thecommercialized form of pioglitazone hydrochloride is a racemic mixtureand adverse side effects have been reported in patients receiving thistherapeutic, including, for example, edema and increased incidence ofbone fracture.

Pioglitazone and other thiazolidinediones have been shown to haveanti-inflammatory activity, part of which seems to be mediated by amechanism not involving PPARs (Curr Drug Targets Inflamm Allergy 2002, 1(3):243-248). Recently, thiazolidinediones have also been shown to bindmitochondrial membrane proteins, including the mitochondrial target ofthiazolidinedione (mTOT), and the thiazolidinediones may modulatemitochondrial metabolism through this direct binding. See, for example,PLoS One. 2013; 8 (5): e61551; PNAS 2013, 110 (14), 5422-5427; Am JPhysiol Endocrinol Metab 2004, 286, E252-260.

Due to the increasing number of patients suffering from disorders suchas those mentioned above, and the limitations of existing therapies,such as adverse side effects, there is a need for new therapeutic agentsfor treating medical disorders in which modulation of PPAR,anti-inflammatory, and/or mTOT activity are predicted to be beneficial.The present invention addresses these needs and provides other relatedadvantages.

SUMMARY OF THE INVENTION

The invention provides enantiopure deuterium-enriched pioglitazone,pharmaceutical compositions, and methods of treating neurologicaldisorders, cancer, respiratory disorders, metabolic disorders, and otherdisorders using the enantiopure deuterium-enriched pioglitazone. Thedeuterated pioglitazone contains deuterium enrichment at the chiralcenter of pioglitazone and optionally in other locations in thecompound. Further, the deuterium-enriched pioglitazone is provided inenantiomerically pure form. This enantiomerically pure,deuterium-enriched pioglitazone provides for a better therapeutic agentthan non-deuterated pioglitazone and/or racemic mixtures ofdeuterium-enriched pioglitazone.

Accordingly, one aspect of the invention provides a deuterium-enrichedcompound of Formula I for use in the therapeutic methods andpharmaceutical compositions described herein. Desirably, thedeuterium-enriched compound of Formula I has an optical purity of atleast 75% enantiomeric excess. Formula I is represented by:

or a pharmaceutically acceptable salt thereof, wherein:

A¹, A², A³, and A⁴ are independently —C(R⁹)(R¹⁰)—;

A⁵ is —C(R¹¹)(R¹²)(R¹³);

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently H or D;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ each represent independently for eachoccurrence H or D; and

Z is H or D, provided that the abundance of deuterium in Z is at least30%.

In certain embodiments, the deuterium-enriched compound used in thetherapeutic methods and pharmaceutical compositions has the followingstructure:

or is a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 90% enantiomeric excess.

Another aspect of the invention provides a deuterium-enriched compoundof Formula II for use in the therapeutic methods and pharmaceuticalcompositions described herein. Desirably, the deuterium-enrichedcompound of Formula II has an optical purity of at least 75%enantiomeric excess. Formula II is represented by:

or a pharmaceutically acceptable salt thereof, wherein:

A¹, A², A³, and A⁴ are independently —C(R⁹)(R¹⁰)—;

A⁵ is —C(R¹¹)(R¹²)(R¹³);

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently H or D;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ each represent independently for eachoccurrence H or D; and

Z is H or D, provided that the abundance of deuterium in Z is at least30%.

In certain embodiments, the deuterium-enriched compound used in thetherapeutic methods and pharmaceutical compositions has the followingstructure:

or is a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 90% enantiomeric excess.

The deuterium-enriched compounds are particularly useful in thetreatment of medical disorders. Exemplary medical disorders include, forexample, neurological disorders, cancer, respiratory disorders, andmetabolic disorders. The compounds are typically administered to apatient in the form of a pharmaceutical composition. Particularlypreferred medical disorders include, for example, Alzheimer's diseaseand other forms of cognitive impairment, non-small cell lung cancer,hepatocellular carcinoma, and chronic obstructive pulmonary disease.

Accordingly, one aspect of the invention provides a method of treating aneurological disorder selected from the group consisting of Alzheimer'sdisease, Parkinson's disease, amyotrophic lateral sclerosis,Friedreich's ataxia, autism spectrum disorder, depression, mildcognitive impairment, Down syndrome, neurodegeneration,adrenoleukodystrophy, Huntington's: disease, stroke, traumatic braininjury, substance abuse, spinal cord injury, neuronal injury, majordepression or bipolar disorder comorbid with metabolic syndrome, and aneurological disorder caused by functional mitochondrial impairment. Themethod comprises administering to a patient in need thereof atherapeutically effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or Formula II, havingan optical purity of at least 75% enantiomeric excess to treat thedisorder. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

Another aspect of the invention provides a method of treating a disorderselected from the group consisting of cancer, a metabolic disorder, asymptom of hepatitis, a cardiovascular disease, polycystic ovarysyndrome, and a skin defect caused by exposure to ultraviolet radiation.The method comprises administering to a patient in need thereof atherapeutically effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or Formula II, havingan optical purity of at least 75% enantiomeric excess to treat thedisorder. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II. In certainembodiments, the disorder is cancer. In certain embodiments, the canceris lung cancer, hepatocellular carcinoma, astrocytoma, glioma,glioblastoma, meningioma, liver cancer, lymphoma, melanoma, multiplemyeloma, pancreatic cancer, colorectal cancer, pituitary cancer, thyroidcancer, esophageal cancer, prostate cancer, ear cancer, nose cancer,throat cancer, kidney cancer, breast cancer, stomach cancer, or uterinecancer. In certain embodiments, the cancer is lung cancer,hepatocellular carcinoma, astrocytoma, glioma, glioblastoma, meningioma,liver cancer, lymphoma, melanoma, multiple myeloma, pancreatic cancer,colorectal cancer, pituitary cancer, thyroid cancer, esophageal cancer,or prostate cancer. In certain embodiments, the cancer is non-small celllung cancer or hepatocellular carcinoma. In certain embodiments, thedisorder is a metabolic disorder, non-alcoholic fatty liver disease,viral hepatitis, liver cirrhosis, liver fibrosis, diabetic retinopathy,diabetic neuropathy, diabetic nephropathy, beta cell depletion, insulinresistance in a patient with congenital adrenal hyperplasia treated witha glucocorticoid, dysmetabolism in peritoneal dialysis patients, reducedinsulin secretion, improper distribution of brown fat cells and whitefat cells, obesity, or improper modulation of leptin levels. In certainembodiments, the disorder is a metabolic disorder selected from thegroup consisting of non-alcoholic fatty liver disease, diabeticretinopathy, diabetic neuropathy, diabetic nephropathy, beta celldepletion, insulin resistance in a patient with congenital adrenalhyperplasia treated with a glucocorticoid, dysmetabolism in peritonealdialysis patients, reduced insulin secretion, improper distribution ofbrown fat cells and white fat cells, obesity, and improper modulation ofleptin levels. In certain embodiments, the disorder is cardiovasculardisease, such as hypertension, hyperlipidemia, atherosclerosis, impropervascular function, dyslipidemia, stenosis, restenosis, myocardialinfarction, stroke, intracranial hemorrhage, acute coronary syndrome,stable angina pectoris, or unstable angina pectoris. In certain otherembodiments, the cardiovascular disorder is intracranial hemorrhage,acute coronary syndrome, stable angina pectoris, or unstable anginapectoris. In certain other embodiments, the metabolic disorder is betacell loss treatable by B-cell regeneration.

Another aspect of the invention provides a method of treating arespiratory disorder. The method comprises administering to a patient inneed thereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat the disorder. In certain embodiments, the deuterium-enrichedcompound is administered by pulmonary administration, such as when thedeuterium-enriched compound has the (S)-stereochemical configuration atthe stereocenter of the thiazolidine-2,4-dione ring. In certainembodiments, the deuterium-enriched compound is administered by routesother than pulmonary administration. In certain embodiments, thedeuterium-enriched compound is administered by oral administration,sublingual administration, sublabial administration, rectaladministration, injection, or transdermal administration. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II. In certain embodiments, the respiratory disorderis further selected from an inflammatory condition that contributes tometabolic syndrome. In certain embodiments, the respiratory disorder ischronic obstructive pulmonary disease, asthma, bronchitis, cysticfibrosis, pulmonary edema, pulmonary embolism, pulmonary arterialhypertension, pneumonia, pulmonary sarcoidosis, silicosis, pulmonaryfibrosis, respiratory failure, acute respiratory distress syndrome,emphysema, chronic bronchitis, tuberculosis, lung cancer, or a chronicrespiratory condition. In certain embodiments, the deuterium-enrichedcompound is administered by oral administration.

Another aspect of the invention provides a method of achieving an effectselected from the group consisting of: (a) reducing the amount of atriglyceride or low-density lipoprotein (LDL) in a patient, and (b)increasing the amount of high-density lipoprotein (HDL) in a patient.The method comprises administering to a patient in need thereof aneffective amount of a deuterium-enriched compound described herein, suchas a compound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to achieve said effect. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II.

Another aspect of the invention provides a method of treating aninflammatory or immune-mediated disorder selected from the groupconsisting chronic kidney disease, arthritis, a primary cicatricialalopecia, lung fibrosis, multiple sclerosis, endotoxemia, sepsis, septicshock, laminitis, inflammatory bowel disease, colitis, Crohn's disease,rheumatoid arthritis, lupus, myasthenia gravis, vasculitis, chronicpancreatitis, a hyperproliferative skin disorder, an inflammatory skindisorder, rhinitis, and a dermatological condition. The method comprisesadministering to a patient in need thereof a therapeutically effectiveamount of a deuterium-enriched compound described herein, such as acompound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to treat the disorder. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II. In yet other embodiments, the inflammatory orimmune-mediated disorder is further selected from an inflammatorycondition that contributes to metabolic syndrome.

Another aspect of the invention provides a method of treating adermatological disorder selected from the group consisting of psoriasis,atopic dermatitis, acne, leukoplakia, scleroderma, and a skinmalignancy. The method comprises administering to a patient in needthereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat the disorder. In certain embodiments, the deuterium-enrichedcompound is a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

Another aspect of the invention provides a method of achieving an effectselected from the group consisting of: (a) modulating expression of apro-inflammatory cytokine (e.g., TNFα, IL-1β, or IL-6) in a patientsuffering from an inflammatory disorder, (b) modulating expression of ananti-inflammatory cytokine in a patient suffering from an inflammatorydisorder, (c) modulating macrophage function in a patient suffering froman infection, and (d) modulating stem cell differentiation in a patient.The method comprises administering to a patient in need thereof aneffective amount of a deuterium-enriched compound described herein, suchas a compound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to achieve said effect. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II.

Another aspect of the invention provides a method of promoting woundhealing. The method comprises administering to a patient in need thereofa therapeutically effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or Formula II, havingan optical purity of at least 75% enantiomeric excess to promote woundhealing. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

Another aspect of the invention provides a method of treating a disorderselected from the group consisting of transplant rejection, liverfunctional impairment, Rabson-Mendenhall syndrome, Donohue syndrome,Leber hereditary optic neuropathy, myotonic dystrophy, ototoxicity,Niemann Pick disease, autosomal dominant optic atrophy, spinal bulbarmuscular atrophy, Mohr-Tranebjaerg syndrome, hereditary spasticparaplegia, MELAS syndrome, monoclonal immunoglobulin deposition disease(MIDD), deafness, insulin resistance in a patient receiving growthhormone, and chronic progressive external ophthalmoplegia withmitochondrial myopathy. The method comprises administering to a patientin need thereof a therapeutically effective amount of adeuterium-enriched compound described herein, such as a compound ofFormula I or Formula II, having an optical purity of at least 75%enantiomeric excess to treat the disorder. In certain embodiments, thedeuterium-enriched compound is a compound of Formula I. In certain otherembodiments, the deuterium-enriched compound is a compound of FormulaII.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a line graph depicting results of PPARγ agonist activitytesting for d-S-pio, d-R-pio, and h-rac-pio, as further described inExample 3.

FIG. 2A is a line graph depicting in vitro stability data for h-S-pio inhuman plasma in the form of experimental data points and results fromfitting to kinetic differential equations, as further described inExample 4. The abbreviation “calc” indicates results from fittingexperimental data to kinetic differential equations.

FIG. 2B is a line graph depicting in vitro stability data for h-R-pio inhuman plasma in the form of experimental data points and results fromfitting to kinetic differential equations, as further described inExample 4. The abbreviation “calc” indicates results from fittingexperimental data to kinetic differential equations.

FIG. 2C is a line graph depicting in vitro stability data for d-rac-pioin human plasma in the form of experimental data points and results fromfitting to kinetic differential equations, as further described inExample 4. The abbreviation “calc” indicates results from fittingexperimental data to kinetic differential equations.

FIG. 3A is a line graph depicting in vitro stability data for h-S-pio inmouse plasma in the form of experimental data points and results fromfitting to kinetic differential equations, as further described inExample 4. The abbreviation “calc” indicates results from fittingexperimental data to kinetic differential equations.

FIG. 3B is a line graph depicting in vitro stability data for h-R-pio inmouse plasma in the form of experimental data points and results fromfitting to kinetic differential equations, as further described inExample 4. The abbreviation “calc” indicates results from fittingexperimental data to kinetic differential equations.

FIG. 3C is a line graph depicting in vitro stability data for d-rac-pioin mouse plasma in the form of experimental data points and results fromfitting to kinetic differential equations, as further described inExample 4. The abbreviation “calc” indicates results from fittingexperimental data to kinetic differential equations.

FIG. 4 is a bar graph depicting maximal respiration as oxygenconsumption rate (OCR in pmoles O₂/min) of C2C12 cells treated withh-rac-pio, d-S-pio, or d-R-pio at 30 μM for 15, 30, or 90 min comparedto the OCR in vehicle-treated cells, as further described in Example 6.

FIG. 5A is a line graph depicting PK profiles for the enantiomers ofpioglitazone in mice administered h-rac-pio (30 mg/kg) by oral gavagedaily for 5 days; (S)-enantiomer—hollow triangles, dashed line;(R)-enantiomer—hollow squares, dotted line; as further described inExample 7.

FIG. 5B is a line graph depicting PK profiles for the enantiomers ofpioglitazone in mice administered d-R-pio (15 mg/kg) by oral gavagedaily for 5 days; (S)-enantiomer—hollow triangles, dashed line;(R)-enantiomer—hollow squares, dotted line; each curve represents thesum of corresponding isotopomers ((S)-enantiomer: h-S-pio+d-S-pio, and(R)-enantiomer: h-R-pio+d-R-pio), as further described in Example 7.

FIG. 5C is a line graph depicting PK profiles for the enantiomers ofpioglitazone in mice administered d-S-pio (15 mg/kg) by oral gavagedaily for 5 days; (S)-enantiomer—hollow triangles, dashed line;(R)-enantiomer—hollow squares, dotted line; each curve represents thesum of corresponding isotopomers ((S)-enantiomer: h-S-pio+d-S-pio, and(R)-enantiomer: h-R-pio+d-R-pio), as further described in Example 7.

FIG. 6 is a line graph depicting the amount of non-fasted blood glucose(ng/mL) in db/db mice dosed daily by oral gavage with vehicle (hollowcircles), 20 mg/kg d-S-pio (filled triangles), 20 mg/kg d-R-pio (filledsquares), or 30 mg/kg h-rac-pio (hollow triangles) measured on day oneat 4 hours pre-first dose, and on days one, eight, and ten, 1 hourpost-dose (2-way repeated measures ANOVA with Sidak post-test,***P<0.001), where the abbreviation “day 1 pre” refers to results frommeasurements taken on day one at 4 hours pre-first dose, and theabbreviation “day 1 post” refers to results from measurements taken onday one at 1 hour post-dose; as further described in Example 8.

FIGS. 7A-C are line graphs depicting the effect of 10-day, daily oraladministration of vehicle, d-S-pio (20 mg/kg), d-R-pio (20 mg/kg), orh-rac-pio (30 mg/kg) to db/db mice on certain metabolic diseasebiomarkers (i.e., insulin, cholesterol, triglycerides, non-essentialfatty acids), adiponectin, and inflammatory biomarkers (i.e., IL-1β,IL-6, TNF-α, MCP-1, and serum amyloid A) (Kruskal-Wallis test withDunn's post-test against vehicle, *P <0.05, **P<0.01, ***P<0.001, and****P<0.0001, for insulin, cholesterol, triglycerides, non-essentialfatty acids, adiponectin, MCP-1; one-tailed unpaired t-test againstvehicle, * P<0.05, for IL-1β, IL-6, TNF-α, and serum amyloid A), asfurther described in Example 8.

FIG. 8 is a line graph depicting the effect of twice daily dosing withvehicle (hollow circles), h-rac-pio (30 mg/kg overall daily dose, hollowtriangles), d-S-pio (15 mg/kg overall daily dose, filled triangles), andd-R-pio (15 mg/kg overall daily dose, filled squares) on body weight,expressed as percent body weight difference versus day 1 body weight(mean±SEM) in male C57BL/6J mice (n=14 mice in vehicle group, n=12 eachin h-rac-pio, d-S-pio, and d-R-pio groups; ANOVA statistical analysiswith multiple comparison Dunnett's post-test, *P<0.05, **P<0.01,***P<0.001), as further described in Example 9.

DETAILED DESCRIPTION

The invention provides enantiopure deuterium-enriched pioglitazone,pharmaceutical compositions, and methods of treating neurologicaldisorders, cancer, respiratory disorders, metabolic disorders, and otherdisorders using enantiopure deuterium-enriched pioglitazone.Deuterium-enriched refers to the feature that the compound has aquantity of deuterium that is greater than in naturally occurringcompounds or synthetic compounds prepared from substrates having thenaturally occurring distribution of isotopes. The threshold amount ofdeuterium enrichment is specified in certain instances in thisdisclosure, and all percentages given for the amount of deuteriumpresent are mole percentages.

Deuterium (²H) is a stable, non-radioactive isotope of ¹H hydrogen andhas an atomic weight of 2.014. Hydrogen naturally occurs as a mixture ofthe isotopes ¹H hydrogen (i.e., protium), deuterium (²H), and tritium(³H). The natural abundance of deuterium is 0.015%. One of ordinaryskill in the art recognizes that in all chemical compounds with an Hatom, the H atom actually represents a mixture of ¹H hydrogen, deuterium(²H), and tritium (³H), where about 0.015% is deuterium. Thus, compoundswith a level of deuterium that has been enriched to be greater than itsnatural abundance of 0.015% are considered unnatural and, as a result,novel over their non-enriched counterparts.

The deuterium-enriched pioglitazone described herein contains deuteriumenrichment at the chiral center of pioglitazone and optionally in otherlocations in the compound. Deuterium-enrichment at the chiral centerreduces the rate at which the two enantiomers of pioglitazone mayinterconvert. Further, the deuterium-enriched pioglitazone describedherein is provided in enantiomerically pure form. This enantiomericallypure, deuterium-enriched pioglitazone provides for a better therapeuticagent than non-deuterated pioglitazone and/or racemic mixtures of thecompound.

Exemplary compositions and methods of the present invention aredescribed in more detail in the following sections: I.Deuterium-enriched Pioglitazone; II. Therapeutic Applications; III.Dosing Considerations and Combination Therapy, and IV. PharmaceuticalCompositions. Aspects of the invention described in one particularsection are not to be limited to any particular section.

I. Deuterium-Enriched Pioglitazone

One aspect of the invention provides deuterium-enriched compounds foruse in the therapeutic methods and pharmaceutical compositions describedherein. The deuterium-enriched compounds are provided in highenantiomeric purity in order to maximize therapeutic benefit, such asmaximal potency per dose of therapeutic agent and minimize adverse sideeffects.

One such deuterium-enriched compound is a family of deuterium-enrichedcompounds represented by Formula I having an optical purity of at least75% enantiomeric excess:

or a pharmaceutically acceptable salt thereof, wherein:

A¹, A², A³, and A⁴ are independently —C(R⁹)(R¹⁰)—;

A⁵ is —C(R¹¹)(R¹²)(R¹³);

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently H or D;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ each represent independently for eachoccurrence H or D; and

Z is H or D, provided that the abundance of deuterium in Z is at least30%.

In certain embodiments, A¹ is —CH₂—. In certain embodiments, A² is—CH₂—. In certain embodiments, A³ is —CH₂—. In certain embodiments, A⁴is —CH₂—. In certain embodiments, A² and A³ are —CH₂—. In certain otherembodiments, A¹, A², A³, and A⁴ are —CH₂—.

In certain embodiments, A⁵ is —CH₃. In certain embodiments, A⁴ is —CH₂—,and A⁵ is —CH₃.

In certain embodiments, R¹ is H. In certain embodiments, R² is H. Incertain embodiments, R³ is H. In certain embodiments, R⁴ is H. Incertain embodiments, R⁵ is H. In certain embodiments, R⁶ is H. Incertain embodiments, R⁷ is H. In certain embodiments, R⁸ is H. Incertain other embodiments, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H.

The description above describes multiple embodiments relating tocompounds of Formula I. The patent application specifically contemplatesall combinations of the embodiments. For example, the inventioncontemplates a compound of Formula I wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷,and R⁸ are H; A¹, A², A³, and A⁴ are —CH₂—; and A⁵ is —CH₃.

Another such deuterium-enriched compound is a family ofdeuterium-enriched compounds represented by Formula I-A having anoptical purity of at least 75% enantiomeric excess:

or a pharmaceutically acceptable salt thereof, wherein Z is H or D,provided that the abundance of deuterium in Z is at least 30%.

The compounds of Formula I and Formula I-A can be further characterizedaccording to the abundance of deuterium at the position defined byvariable Z. In certain embodiments, the abundance of deuterium in Z isselected from: (a) at least 40%, (b) at least 50%, (c) at least 60%, (d)at least 70%, (e) at least 75%, (f) at least 80%, (g) at least 90%, (h)at least 95%, (h) at least 97%, and (i) about 100%. Additional examplesof the abundance of deuterium in Z include 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99 to about 100%.

In certain embodiments, the abundance of deuterium in Z is at least 60%.In certain other embodiments, the abundance of deuterium in Z is atleast 75%. In yet other embodiments, the abundance of deuterium in Z isat least 90%.

The compounds of Formula I and Formula I-A can be further characterizedaccording their enantiomeric purity. In certain embodiments, thedeuterium-enriched compound has an enantiomeric excess of at least 80%,85%, 90%, 95%, or 98%. Still further examples of the optical purityinclude an enantiomeric excess of at least 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, or 99%.

Still further such deuterium-enriched compounds are provided in Tables 1and 2 below.

TABLE 1 Compound No. Structure  1

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 60% enantiomeric excess.  2

  having an optical purity of at least 60% enantiomeric excess.  3

  hydrochloride having an optical purity of at least 60% enantiomericexcess.  4

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 75% enantiomeric excess.  5

  having an optical purity of at least 75% enantiomeric excess.  6

  hydrochloride having an optical purity of at least 75% enantiomericexcess.  7

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 90% enantiomeric excess.  8

  having an optical purity of at least 90% enantiomeric excess.  9

  hydrochloride having an optical purity of at least 90% enantiomericexcess. 10

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 95% enantiomeric excess. 11

  having an optical purity of at least 95% enantiomeric excess. 12

  hydrochloride having an optical purity of at least 95% enantiomericexcess.

TABLE 2

Compound No. Variable Definition 1 Z = D; R¹ = D; R²-R⁸ are H; A¹, A²,A³, and A⁴ are —CH₂—; and A⁵ is —CH₃ 2 Z = D; R¹-R⁸ are H; A¹ = —CD₂—;A², A³, and A⁴ are —CH₂—; and A⁵ is —CH₃ 3 Z = D; R¹ = H; R², R³, R⁴,and R⁵ are D; R⁶-R⁸ are H; A¹, A², A³, and A⁴ are —CH₂—; and A⁵ is —CH₃4 Z = D; R¹-R⁸ are H; A¹ = —CH₂—; A² and A³ are —CD₂—; A⁴ = —CH₂—; andA⁵ is —CH₃ 5 Z = D; R¹, R², R³, R⁴, and R⁵ are H; R⁶-R⁸ are D; A¹, A²,A³, and A⁴ are —CH₂—; and A⁵ is —CH₃ 6 Z = D; R¹-R⁸ are H; A¹, A², andA³ are —CH₂—; A⁴ are —CD₂—; and A⁵ is —CD₃

Another embodiment of the invention provides a compound in Table 2wherein the compound has an enantiomeric excess of at least 60%, 70%,75%, 80%, 85%, 90%, 95%, or 98%.

Another such deuterium-enriched compound is a family ofdeuterium-enriched compounds represented by Formula II having an opticalpurity of at least 75% enantiomeric excess:

or a pharmaceutically acceptable salt thereof, wherein:

A¹, A², A³, and A⁴ are independently —C(R⁹)(R¹⁰)—;

A⁵ is —C(R¹¹)(R¹²)(R¹³);

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently H or D;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ each represent independently for eachoccurrence H or D; and

Z is H or D, provided that the abundance of deuterium in Z is at least30%.

In certain embodiments, A¹ is —CH₂—. In certain embodiments, A² is—CH₂—. In certain embodiments, A³ is —CH₂—. In certain embodiments, A⁴is —CH₂—. In certain embodiments, A² and A³ are —CH₂-. In certain otherembodiments, A¹, A², A³, and A⁴ are —CH₂—.

In certain embodiments, A⁵ is —CH₃. In certain embodiments, A⁴ is —CH₂—,and A⁵ is —CH₃.

In certain embodiments, R¹ is H. In certain embodiments, R² is H. Incertain embodiments, R³ is H. In certain embodiments, R⁴ is H. Incertain embodiments, R⁵ is H. In certain embodiments, R⁶ is H. Incertain embodiments, R⁷ is H. In certain embodiments, R⁸ is H. Incertain other embodiments, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H.

The description above describes multiple embodiments relating tocompounds of Formula II. The patent application specificallycontemplates all combinations of the embodiments. For example, theinvention contemplates a compound of Formula II wherein R¹, R², R³, R⁴,R⁵, R⁶, R⁷, and R⁸ are H; A¹, A², A³, and A⁴ are —CH₂—; and A⁵ is —CH₃.

Another such deuterium-enriched compound is a family ofdeuterium-enriched compounds represented by Formula II-A having anoptical purity of at least 75% enantiomeric excess:

or a pharmaceutically acceptable salt thereof, wherein Z is H or D,provided that the abundance of deuterium in Z is at least 30%.

The compounds of Formula II and Formula II-A can be furthercharacterized according to the abundance of deuterium at the positiondefined by variable Z. In certain embodiments, the abundance ofdeuterium in Z is selected from: (a) at least 40%, (b) at least 50%, (c)at least 60%, (d) at least 70%, (e) at least 75%, (f) at least 80%, (g)at least 90%, (h) at least 95%, (h) at least 97%, and (i) about 100%.Additional examples of the abundance of deuterium in Z include 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 to about 100%.

In certain embodiments, the abundance of deuterium in Z is at least 60%.In certain other embodiments, the abundance of deuterium in Z is atleast 75%. In yet other embodiments, the abundance of deuterium in Z isat least 90%.

The compounds of Formula II and Formula II-A can be furthercharacterized according their enantiomeric purity. In certainembodiments, the deuterium-enriched compound has an enantiomeric excessof at least 80%, 85%, 90%, 95%, or 98%. Still further examples of theoptical purity include an enantiomeric excess of at least 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, or 99%.

Still further such deuterium-enriched compounds are provided in Tables 3and 4 below.

TABLE 3 Compound No. Structure  1

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 60% enantiomeric excess.  2

  having an optical purity of at least 60% enantiomeric excess.  3

  hydrochloride having an optical purity of at least 60% enantiomericexcess.  4

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 75% enantiomeric excess.  5

  having an optical purity of at least 75% enantiomeric excess.  6

  hydrochloride having an optical purity of at least 75% enantiomericexcess.  7

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 90% enantiomeric excess.  8

  having an optical purity of at least 90% enantiomeric excess.  9

  hydrochloride having an optical purity of at least 90% enantiomericexcess. 10

  or a pharmaceutically acceptable salt thereof, each having an opticalpurity of at least 95% enantiomeric excess. 11

  having an optical purity of at least 95% enantiomeric excess. 12

  hydrochloride having an optical purity of at least 95% enantiomericexcess.

TABLE 4

Compound No. Variable Definition 1 Z = D; R¹ = D; R²-R⁸ are H; A¹, A²,A³, and A⁴ are —CH₂—; and A⁵ is —CH₃ 2 Z = D; R¹-R⁸ are H; A¹ = —CD₂—;A², A³, and A⁴ are —CH₂—; and A⁵ is —CH₃ 3 Z = D; R¹ = H; R², R³, R⁴,and R⁵ are D; R⁶-R⁸ are H; A¹, A², A³, and A⁴ are —CH₂—; and A⁵ is —CH₃4 Z = D; R¹-R⁸ are H; A¹ = —CH₂—; A² and A³ are —CD₂—; A⁴ = —CH₂—; andA⁵ is —CH₃ 5 Z = D; R¹, R², R³, R⁴, and R⁵ are H; R⁶-R⁸ are D; A¹, A²,A³, and A⁴ are —CH₂—; and A⁵ is —CH₃ 6 Z = D; R¹-R⁸ are H; A¹, A², andA³ are —CH₂—; A⁴ are —CD₂—; and A⁵ is —CD₃

Another embodiment of the invention provides a compound in Table 4wherein the compound has an enantiomeric excess of at least 60%, 70%,75%, 80%, 85%, 90%, 95%, or 98%.

As indicated above, the deuterium-enriched compound may be in the formof a pharmaceutically acceptable salt. One such pharmaceuticallyacceptable salt is a hydrochloride salt.

It is understood that the deuterium-enriched compounds described hereincan be combined with a pharmaceutically acceptable carrier to form apharmaceutical composition.

Deuterium-enriched compounds of the invention can generally be preparedby substituting a deuterium-enriched reagent for a non-isotopicallylabeled reagent in synthetic schemes reported in the literature formaking non-isotopically labeled pioglitazone. Scheme 1 below illustratesa general method for preparing deuterium-enriched pioglitazone, in whichdeuterium is incorporated at the sole chiral center. The scheme isprovided for the purpose of illustrating the invention, and should notbe regarded in any manner as limiting the scope or the spirit of theinvention. In Scheme 1, pioglitazone hydrochloride is first stirred withperdeuterated dimethylsulfoxide (d₆-DMSO) and triethylamine and thentreated with perdeuterated methanol (d₄-MeOH). The R-enantiomer andS-enantiomer of deutero-amine A are separated using chiralchromatography, such as chiral high-performance liquid chromatography.Alternatively, the R-enantiomer and S-enantiomer of deutero-amine A maybe separated by reaction with a chiral carboxylic acid to form a salt,followed by separation of the resulting diastereomeric salts, andconversion of the separated salts back to deuterated pioglitazone freebase in enantio-pure form. Pioglitazone hydrochloride can be preparedusing the methods described in for example, U.S. Pat. No. 4,444,779; EP193256; U.S. Pat. Nos. 4,687,777; 8,173,816; and U.S. Patent ApplicationPublication No. 2011/0021576, each of which is incorporated herein byreference.

Scheme 2 below illustrates a general method for preparingdeuterium-enriched pioglitazone, in which deuterium is incorporated atthe ethyl group attached to the pyridine and at the sole chiral center.Reaction of 2-(5-(d₅-ethyl)pyridin-2-yl)ethanol (A1) withp-fluoro-nitrobenzene provides nitrophenyl ether B1. Reduction ofnitrophenyl ether B1, such as through hydrogenation in the presence ofpalladium/carbon, provides aminophenyl ether C1. Reaction of aminophenylether C1 with NaNO₂ and hydrobromic acid, followed by addition ofCH₂═CHCO₂Et provides alpha-bromo ester D1. Reaction of alpha-bromo esterD1 with thiourea provides thiazolidine-2,4-dione E1. Reaction ofthiazolidine-2,4-dione E1 with d₆-DMSO and triethylamine, followed byd₄-MeOH provides deutero-thiazolidine-2,4-dione F1. The R-enantiomer andS-enantiomer of deutero-thiazolidine-2,4-dione F1 are separated usingchiral chromatography, such as chiral high-performance liquidchromatography.

Compounds having deuterium enrichment at a position other than the ethylgroup on the pyridine can be prepared using other deuterated forms ofstarting materials shown in Scheme 2 (e.g., a deuterated form ofp-fluoro-nitrobenzene).

Compounds described herein can be provided in isolated or purified form.Isolated or purified compounds are a group of compounds that have beenseparated from their environment, such as from a crude reaction mixtureif made in a laboratory setting or removed from their naturalenvironment if naturally occurring. Examples of the purity of theisolated compound include, for example, at least 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, to 100% by weight.

Another aspect of the invention provides a unit quantum of adeuterium-enriched compound described herein, such as an amount of atleast (a) one μg of a disclosed deuterium-enriched compound, (b) one mg,or (c) one gram. In further embodiments, the quantum is, for example, atleast 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, or 1 moleof the compound. The present amounts also cover lab-scale (e.g., gramscale including 1, 2, 3, 4, 5 g, etc.), kilo-lab scale (e.g., kilogramscale including 1, 2, 3, 4, 5 kg, etc.), and industrial or commercialscale (e.g., multi-kilogram or above scale including 100, 200, 300, 400,500 kg, etc.) quantities as these will be more useful in the actualmanufacture of a pharmaceutical. Industrial/commercial scale refers tothe amount of product that would be produced in a batch that wasdesigned for clinical testing, formulation, sale/distribution to thepublic, etc.

II. Therapeutic Applications

The invention provides methods of using deuterium-enriched compoundsdescribed herein to treat medical disorders. Preferred medical disordersfor treatment include neurological disorders, cancer, respiratorydisorders, and metabolic disorders. Use of the deuterium-enrichedcompounds having high enantiomeric purity is contemplated to maximizetherapeutic benefit, such as achieving increased potency per dose oftherapeutic agent and minimize adverse side effects. Thedeuterium-enriched compound can be, for example, a compound of FormulaI, Formula I-A, Formula II, Formula II-A, or one of the otherdeuterium-enriched compounds described in Section I above.

Treating Neurological Disorders

Accordingly, one aspect of the invention provides a method of treating aneurological disorder selected from the group consisting of Alzheimer'sdisease, Parkinson's disease, amyotrophic lateral sclerosis,Friedreich's ataxia, autism spectrum disorder, depression, mildcognitive impairment, Down syndrome, neurodegeneration,adrenoleukodystrophy, Huntington's disease, stroke, traumatic braininjury, substance abuse, spinal cord injury, neuronal injury, majordepression or bipolar disorder comorbid with metabolic syndrome, and aneurological disorder caused by functional mitochondrial impairment. Themethod comprises administering to a patient in need thereof atherapeutically effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or Formula II, havingan optical purity of at least 75% enantiomeric excess to treat thedisorder. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II. In certainembodiments, the neurological disorder is selected from the groupconsisting of Alzheimer's disease, Parkinson's disease, amyotrophiclateral sclerosis, Friedreich's ataxia, autism spectrum disorder,depression, mild cognitive impairment, neurodegeneration,adrenoleukodystrophy, Huntington's disease, stroke, traumatic braininjury, substance abuse, spinal cord injury, neuronal injury, and majordepression or bipolar disorder comorbid with metabolic syndrome. Incertain embodiments, the neurological disorder is selected from thegroup consisting of Alzheimer's disease, Parkinson's disease,amyotrophic lateral sclerosis, Friedreich's ataxia, depression, mildcognitive impairment, neurodegeneration, adrenoleukodystrophy, andHuntington's disease. In certain other embodiments, the neurologicaldisorder is Alzheimer's disease. In certain other embodiments, theneurological disorder is Down syndrome.

In certain other embodiments, the neurological disorder is a cognitivedisorder, such as cognitive impairment and/or memory impairment. Thecognitive impairment may be, for example, cognitive impairmentassociated with Alzheimer's disease.

In certain embodiments, the substance abuse is one or more of alcoholcraving, heroin dependence, and nicotine dependence.

Treating Cancer

Another aspect of the invention provides a method of treating cancer.The method comprises administering to a patient in need thereof atherapeutically effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or Formula II, havingan optical purity of at least 75% enantiomeric excess to treat thecancer. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

In certain embodiments, the cancer is lung cancer, hepatocellularcarcinoma, astrocytoma, glioma, glioblastoma, meningioma, liver cancer,lymphoma, melanoma, multiple myelorna, pancreatic cancer, colorectalcancer, pituitary cancer, thyroid cancer, esophageal cancer, or prostatecancer. In certain embodiments, the cancer is non-small cell lung canceror hepatocellular carcinoma.

In certain other embodiments, the cancer is lung cancer, hepatocellularcarcinoma, astrocytoma, glioma, glioblastoma, meningioma, liver cancer,lymphoma, melanoma, multiple myeloma, pancreatic cancer, colorectalcancer, pituitary cancer, thyroid cancer, esophageal cancer, prostatecancer, ear cancer, nose cancer, throat cancer, kidney cancer, breastcancer, stomach cancer, or uterine cancer. In certain other embodiments,the cancer is brain cancer, bladder cancer, breast cancer, cervicalcancer, colon cancer, colorectal cancer, endometrial cancer, esophagealcancer, leukemia, lung cancer, liver cancer, melanoma, ovarian cancer,pancreatic cancer, prostate cancer, rectal cancer, renal cancer, stomachcancer, testicular cancer, or uterine cancer. In yet other embodiments,the cancer is a vascularized tumor, squamous cell carcinoma,adenocarcinoma, small cell carcinoma, melanoma, glioma, neuroblastoma,sarcoma (e.g., an angiosarcoma or chondrosarcoma), larynx cancer,parotid cancer, biliary tract cancer, thyroid cancer, acral lentiginousmelanoma, actinic keratosis, acute lymphocytic leukemia, acute myeloidleukemia, adenoid cystic carcinoma, adenoma, adenosarcoma, adenosquamouscarcinoma, anal canal cancer, anal cancer, anorectal cancer, astrocytictumor, Bartholin's gland carcinoma, basal cell carcinoma, biliarycancer, bone cancer, bone marrow cancer, bronchial cancer, bronchialgland carcinoma, carcinoid, cholangiocarcinoma, chorioid plexuspapilloma/carcinoma, chronic lymphocytic leukemia, chronic myeloidleukemia, clear cell carcinoma, connective tissue cancer, cystadenoma,digestive system cancer, duodenum cancer, endocrine system cancer,endodermal sinus tumor, endometrial hyperplasia, endometrial stromalsarcoma, endometrioid adenocarcinoma, endothelial cell cancer, ependymalcancer, epithelial cell cancer, Ewing's sarcoma, eye and orbit cancer,female genital cancer, focal nodular hyperplasia, gallbladder cancer,gastric antrum cancer, gastric fundus cancer, gastrinoma, glioblastoma,glucagonoma, heart cancer, hemangioblastoma, hemangioendothelioma,hemangioma, hepatic adenoma, hepatic adenomatosis, hepatobiliary cancer,hepatocellular carcinoma, Hodgkin's disease, ileum cancer, insulinoma,intraepithelial neoplasia, interepithelial squamous cell neoplasia,intrahepatic bile duct cancer, invasive squamous cell carcinoma, jejunumcancer, joint cancer, Kaposi's sarcoma, pelvic cancer, large cellcarcinoma, large intestine cancer, leiomyosarcoma, lentigo malignamelanoma, lymphoma, male genital cancer, malignant melanoma, malignantmesothelial tumor, medulloblastoma, medulloepithelioma, meningealcancer, mesothelial cancer, metastatic carcinoma, mouth cancer,mucoepidermoid carcinoma, multiple myeloma, muscle cancer, nasal tractcancer, nervous system cancer, neuroepithelial adenocarcinoma, nodularmelanoma, non-epithelial skin cancer, non-Hodgkin's lymphoma, oat cellcarcinoma, oligodendroglial cancer, oral cavity cancer, osteosarcoma,papillary serous adenocarcinoma, penile cancer, pharynx cancer,pituitary tumor, plasmacytoma, pseudosarcoma, pulmonary blastoma, rectalcancer, renal cell carcinoma, respiratory system cancer, retinoblastoma,rhabdomyosarcoma, sarcoma, serous carcinoma, sinus cancer, skin cancer,small cell carcinoma, small intestine cancer, smooth muscle cancer, softtissue cancer, somatostatin-secreting tumor, spine cancer, squamous cellcarcinoma, striated muscle cancer, submesothelial cancer, superficialspreading melanoma, T-cell leukemia, tongue cancer, undifferentiatedcarcinoma, ureter cancer, urethral cancer, urinary bladder cancer,urinary system cancer, uterine cervix cancer, uterine corpus cancer,uveal melanoma, vaginal cancer, verrucous carcinoma, VIPoma, vulvacancer, well differentiated carcinoma, or Wilms tumor.

In certain other embodiments, the cancer is non-Hodgkin's lymphoma, suchas a B-cell lymphoma or a T-cell lymphoma. In certain embodiments, thenon-Hodgkin's lymphoma is a B-cell lymphoma, such as a diffuse largeB-cell lymphoma, primary mediastinal B-cell lymphoma, follicularlymphoma, small lymphocytic lymphoma, mantle cell lymphoma, marginalzone B-cell lymphoma, extranodal marginal zone B-cell lymphoma, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma,Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, orprimary central nervous system (CNS) lymphoma. In certain otherembodiments, the non-Hodgkin's lymphoma is a T-cell lymphoma, such as aprecursor T-lymphoblastic lymphoma, peripheral T-cell lymphoma,cutaneous T-cell lymphoma, angioimmunoblastic T-cell lymphoma,extranodal natural killer/T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplasticlarge cell lymphoma, or peripheral T-cell lymphoma.

Treating Respiratory Disorders

Another aspect of the invention provides a method of treating arespiratory disorder. The method comprises administering to a patient inneed thereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat the disorder. In certain embodiments, the deuterium-enrichedcompound is a compound described herein having the (S)-stereochemicalconfiguration at the stereocenter of the thiazolidine-2,4-dione ring. Incertain embodiments, the deuterium-enriched compound is a compound ofFormula I. In certain other embodiments, the deuterium-enriched compoundis a compound of Formula II.

In certain embodiments, the deuterium-enriched compound is administeredby pulmonary administration. In a more specific embodiment, thedeuterium-enriched compound is a compound described herein having the(S)-stereochemical configuration at the stereocenter of thethiazolidine-2,4-dione ring, and said compound is administered bypulmonary administration. In another more specific embodiment, thedeuterium-enriched compound is a compound of Formula I and said compoundis administered by pulmonary administration.

In certain embodiments, the deuterium-enriched compound is administeredby routes other than pulmonary administration. In certain embodiments,the deuterium-enriched compound is administered by oral administration,sublingual administration, sublabial administration, rectaladministration, injection, or transdermal administration. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II.

In certain embodiments, the respiratory disorder is chronic obstructivepulmonary disease, asthma, bronchitis, cystic fibrosis, pulmonary edema,pulmonary embolism, pulmonary arterial hypertension, pneumonia,pulmonary sarcoidosis, silicosis, pulmonary fibrosis, respiratoryfailure, acute respiratory distress syndrome, emphysema, chronicbronchitis, tuberculosis, lung cancer, or a chronic respiratorycondition. In certain embodiments, the respiratory disorder is chronicobstructive pulmonary disease, asthma, or a chronic respiratorycondition. In certain other embodiments, the respiratory disorder ischronic obstructive pulmonary disease. In yet other embodiments, therespiratory disorder is bronchitis, cystic fibrosis, pulmonary edema,pulmonary embolism, pneumonia, pulmonary sarcoidosis, silicosis,pulmonary fibrosis, respiratory failure, acute respiratory distresssyndrome, emphysema, chronic bronchitis, tuberculosis, or lung cancer.In certain embodiments, the asthma is mild asthma, moderate asthma,severe asthma, or steroid-resistant asthma.

Treating Metabolic Disorders

Another aspect of the invention provides a method of treating ametabolic disorder selected from the group consisting of non-alcoholicfatty liver disease, viral hepatitis, liver cirrhosis, liver fibrosis,diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, betacell depletion insulin resistance in a patient with congenital adrenalhyperplasia treated with a glucocorticoid, dysmetabolism in peritonealdialysis patients, reduced insulin secretion, improper distribution ofbrown fat cells and white fat cells, obesity, or improper modulation ofleptin levels. The method comprises administering to a patient in needthereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat the disorder. In certain embodiments, the deuterium-enrichedcompound is a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II. In certainembodiments, the metabolic disorder is further selected from acomplication of diabetes. In certain embodiments, the metabolic disorderis non-alcoholic fatty liver disease, diabetic retinopathy, diabeticneuropathy, diabetic nephropathy, or beta cell depletion insulinresistance in a patient with congenital adrenal hyperplasia treated witha glucocorticoid. In certain embodiments, the metabolic disorder isnon-alcoholic fatty liver disease, diabetic retinopathy, diabeticneuropathy, diabetic nephropathy, beta cell depletion, reduced insulinsecretion, improper distribution of brown fat cells and white fat cells,obesity, or improper modulation of leptin levels. In certain otherembodiments, the metabolic disorder is non-alcoholic fatty liverdisease. In certain other embodiments, the metabolic disorder is betacell loss treatable by B-cell regeneration. In certain otherembodiments, the metabolic disorder is central obesity, dyslipidemia, orpre-diabetes.

Treating a Symptom of Hepatitis

Another aspect of the invention provides a method of treating a symptomof hepatitis. The method comprises administering to a patient in needthereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat a symptom of hepatitis. In certain embodiments, thedeuterium-enriched compound is a compound of Formula I. In certain otherembodiments, the deuterium-enriched compound is a compound of Formula II

Treating Cardiovascular Disease

Another aspect of the invention provides a method of treating acardiovascular disease. The method comprises administering to a patientin need thereof a therapeutically effective amount of adeuterium-enriched compound described herein, such as a compound ofFormula I or Formula II, having an optical purity of at least 75%enantiomeric excess to treat the cardiovascular disease. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II. In certain embodiments, the cardiovasculardisease is hypertension, hyperlipidemia, atherosclerosis, impropervascular function, dyslipidemia, stenosis, restenosis, myocardialinfarction, stroke, intracranial hemorrhage, acute coronary syndrome,stable angina pectoris, or unstable angina pectoris. In certain otherembodiments, the cardiovascular disorder is intracranial hemorrhage,acute coronary syndrome, stable angina pectoris, or unstable anginapectoris.

In another aspect, the invention provides a method for preventing strokein a patient. The method comprises administering to a patient in needthereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess toprevent said stroke.

The method of treatment or the method of prevention may involve apatient at risk for central nervous system ischemic stroke, or mayinvolve a patient at risk for stroke due to cardiovascular disease.

Reducing the Amount of a Triglyceride or Low-Density Lipoprotein

Another aspect of the invention provides a method of reducing the amountof a triglyceride or low-density lipoprotein (LDL) in a patient. Themethod comprises administering to a patient in need thereof an effectiveamount of a deuterium-enriched compound described herein, such as acompound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to reduce the amount of a triglyceride orLDL in the patient. In certain embodiments, the deuterium-enrichedcompound is a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

In certain embodiments, the method provides a reduction of at least 1%,5%, 10%, or 25% in the amount of a triglyceride or low-densitylipoprotein (LDL) in the patient.

Increasing the Amount of High-Density Lipoprotein

Another aspect of the invention provides a method of increasing theamount of high-density lipoprotein (HDL) in a patient. The methodcomprises administering to a patient in need thereof a therapeuticallyeffective amount of a deuterium-enriched compound described herein, suchas a compound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to increase the amount of HDL in thepatient. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

In certain embodiments, the method provides an increase of at least 1%,5%, 10%, or 25% in the amount of high-density lipoprotein (HDL) in apatient.

Treating an Inflammatory or Immune-Mediate Disorder

Another aspect of the invention provides a method of treating aninflammatory or immune-mediated disorder selected from the groupconsisting of chronic kidney disease, arthritis, a primary cicatricialalopecia, lung fibrosis, multiple sclerosis, endotoxemia, sepsis, septicshock, laminitis, inflammatory bowel disease, colitis, Crohn's disease,rheumatoid. arthritis, lupus, myasthenia gravis, vasculitis, chronicpancreatitis, a hyperproliferative skin disorder, an inflammatory skindisorder, rhinitis (e.g., allergic rhinitis), and a dermatologicalcondition. The method comprises administering to a patient in needthereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat the disorder. In certain embodiments, the deuterium-enrichedcompound is a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II. In certainembodiments, the inflammatory or immune-mediated disorder is selectedfrom the group consisting of chronic kidney disease, arthritis, aprimary cicatricial alopecia, lung fibrosis, multiple sclerosis,endotoxemia, sepsis, septic shock, laminitis, inflammatory boweldisease, colitis, Crohn's disease, rheumatoid arthritis, lupus,myasthenia gravis, vasculitis, chronic pancreatitis, ahyperproliferative skin disorder, an inflammatory skin disorder, and adermatological condition. In certain embodiments, the inflammatory orimmune-mediated disorder is selected from the group consisting ofchronic kidney disease, arthritis, a primary cicatricial alopecia, lungfibrosis, multiple sclerosis, endotoxemia, sepsis, septic shock,laminitis, inflammatory bowel disease, colitis, Crohn's disease,rheumatoid arthritis, lupus, myasthenia gravis, vasculitis, chronicpancreatitis, a hyperproliferative skin disorder, an inflammatory skindisorder, and a dermatological condition. In certain embodiments, thechronic kidney disease may be, for example, polycystic kidney disease(such as autosomal dominant or autosomal recessive).

Treating a Dermotological Disorder

Another aspect of the invention provides a method of treating adermatological disorder selected from the group consisting of psoriasis,atopic dermatitis, acne, leukoplakia, scleroderma, and a skinmalignancy. The method comprises administering to a patient in needthereof a therapeutically effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess totreat the disorder. In certain embodiments, the deuterium-enrichedcompound is a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

Modulating Expression of Pro-Inflammatory Cytokines

Another aspect of the invention provides a method of modulatingexpression of a pro-inflammatory cytokine (e.g., TNFα, IL-1β, or IL-6)in a patient suffering from an inflammatory disorder. The methodcomprises administering to a patient in need thereof an effective amountof a deuterium-enriched compound described herein, such as a compound ofFormula I or Formula II, having an optical purity of at least 75%enantiomeric excess to modulate expression of the pro-inflammatorycytokine. In certain embodiments, the pro-inflammatory cytokine is TNFα.In certain embodiments, the deuterium-enriched compound is a compound ofFormula I. In certain other embodiments, the deuterium-enriched compoundis a compound of Formula II.

Another aspect of the invention provides a method of modulatingexpression of an anti-inflammatory cytokine in a patient suffering froman inflammatory disorder. The method comprises administering to apatient in need thereof an effective amount of a deuterium-enrichedcompound described herein, such as a compound of Formula I or FormulaII, having an optical purity of at least 75% enantiomeric excess tomodulate expression of the anti-inflammatory cytokine. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II.

Modulating Macrophage Function

Another aspect of the invention provides a method of modulatingmacrophage function in a patient suffering from an infection. The methodcomprises administering to a patient in need thereof an effective amountof a deuterium-enriched compound described herein, such as a compound ofFormula I or Formula II, having an optical purity of at least 75%enantiomeric excess to modulate macrophage function. In certainembodiments, the deuterium-enriched compound is a compound of Formula I.In certain other embodiments, the deuterium-enriched compound is acompound of Formula II.

Method of Promoting Wound Healing

Another aspect of the invention provides a method of promoting woundhealing. The method comprises administering to a patient in need thereofa therapeutically effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or Formula II, havingan optical purity of at least 75% enantiomeric excess to promote woundhealing. In certain embodiments, the deuterium-enriched compound is acompound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

Treating Skin Defects

Another aspect of the invention provides a method of treating skindefects caused by exposure to ultraviolet radiation. The methodcomprises administering to a patient in need thereof a therapeuticallyeffective amount of a deuterium-enriched compound described herein, suchas a compound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to treat skin defects caused by exposureto ultraviolet radiation. In certain embodiments, the deuterium-enrichedcompound is a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II.

Method of Modulating Stem Cell Differentation

Another aspect of the invention provides a method of modulating stemcell differentiation, such as in a patient. The method comprisesexposing a stem cell to a deuterium-enriched compound described herein,such as a compound of Formula I or Formula II, having an optical purityof at least 75% enantiomeric excess to modulate stem celldifferentiation. In certain embodiments, the deuterium-enriched compoundis a compound of Formula I. In certain other embodiments, thedeuterium-enriched compound is a compound of Formula II. In certainembodiments, the method modulates stem cell differentiation in a patientby administering to the patient an effective amount of a compoundherein, such as a compound of Formula I or Formula II, having an opticalpurity of at least 75% enantiomeric excess.

Additional Medical Disorders

Another aspect of the invention provides a method of treating a disorderselected from the group consisting of transplant rejection, liverfunctional impairment, Rabson-Mendenhall syndrome, Donohue syndrome,Leber hereditary optic neuropathy, myotonic dystrophy, ototoxicity,Niemann Pick disease, autosomal dominant optic atrophy, spinal bulbarmuscular atrophy, Mohr-Tranebjaerg syndrome, hereditary spasticparaplegia, MELAS syndrome, monoclonal immunoglobulin deposition disease(MIDD), deafness, insulin resistance in a patient receiving growthhormone, and chronic progressive external ophthalmoplegia withmitochondrial myopathy. The method comprises administering to a patientin need thereof a therapeutically effective amount of adeuterium-enriched compound described herein, such as a compound ofFormula I or Formula II, having an optical purity of at least 75%enantiomeric excess to treat the disorder. In certain embodiments, thedeuterium-enriched compound is a compound of Formula I. In certain otherembodiments, the deuterium-enriched compound is a compound of FormulaII.

Preventing Medical Disorders

Also provided are methods of preventing a medical disorder in a patient.The method comprises administering to a patient in need thereof aneffective amount of a deuterium-enriched compound described herein, suchas a compound of Formula I or Formula II, having an optical purity of atleast 75% enantiomeric excess to prevent the medical disorder. Themedical disorder may be one or more of the medical disorders recitedabove, such as a neurological disorder (e.g., Alzheimer's disease orParkinson's disease), cancer (e.g., non-small cell lung cancer orhepatocellular carcinoma), a metabolic disorder, a cardiovasculardisorder (e.g. in-stent renarrowing in diabetes patients, reinfarctionin diabetes patients, or cardiac allograft vasculopathy after hearttransplant), or a respiratory disorder (e.g., chronic obstructivepulmonary disease).

Additional Medical Uses

Also provided are methods of using compounds herein for therapycomprising regenerative medicine. Also provided are methods of treatingveterinary disorders, such as laminitis, using a compound describedherein, such as a compound of Formula I or Formula II, having an opticalpurity of at least 75% enantiomeric excess to treat the veterinarydisorder.

Manufacture of Medicaments

Another aspect of the invention provides for the use of adeuterium-enriched compound described herein in the manufacture of amedicament. The medicament may be for treating one or more of themedical disorders described herein, such as treating a neurologicaldisorder (e.g., Alzheimer's disease or Parkinson's disease), cancer(e.g., non-small cell lung cancer or hepatocellular carcinoma), ametabolic disorder, or a respiratory disorder (e.g., chronic obstructivepulmonary disease).

III. Dosing Considerations and Combination Therapy

Doses of a compound provided herein, or a pharmaceutically acceptablesalt thereof, vary depending on factors such as: specific indication tobe treated; age and condition of a patient; and amount of second activeagent used, if any. Generally, a compound provided herein, or apharmaceutically acceptable salt thereof, may be used in an amount offrom about 0.1 mg to about 1 g per day, or from about 0.1 mg to about500 mg per day, and can be adjusted in a conventional fashion (e.g., thesame amount administered each day of the treatment), in cycles (e.g.,one week on, one week oft), or in an amount that increases or decreasesover the course of treatment. In other embodiments, the dose can be fromabout 1 mg to about 500 mg, from about 0.1 mg to about 150 mg, fromabout 1 mg to about 300 mg, from about 10 mg to about 100 mg, from about0.1 mg to about 50 mg, from about 1 mg to about 50 mg, from about 10 mgto about 50 mg, from about 20 mg to about 30 mg, or from about 1 mg toabout 20 mg.

In yet other embodiments, the daily dose can be from about 1 mg to 5 mg,5 mg to 10 mg, 10 mg to 20 mg, 20 mg to 35 mg, 35 mg to 50 mg, 50 mg to75 mg, 75 mg to 100 mg, 100 mg to 125 mg, 125 mg to 150 mg, 150 mg to175 mg, 175 mg to 200 mg, 200 mg to 225 mg, 225 mg to 250 mg, 250 mg to275 mg, 275 mg to 300 mg, 300 mg to 325 mg, 325 mg to 350 mg, 350 mg to375 mg, 375 mg to 400 mg, 400 mg to 425 mg, 425 mg to 450 mg, 450 mg to475 mg, or 475 mg to 500 mg. In certain embodiments, the daily dosage isin the range of about 1 mg to 50 mg, 50 mg to 100 mg, 100 mg to 150 mg,150 mg to 200 mg, 200 mg to 250 mg, 250 mg to 300 mg, 300 mg to 350 mg,350 mg to 400 mg, or 400 mg to 500 mg. In yet other embodiments, thedaily dose is less than about 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400mg, 425 mg, or 450 mg. In yet other embodiments, the daily dose is lessthan about 125 mg, 150 mg, or 175 mg.

Unless indicated otherwise, compounds described herein may beadministered using any medically accepted route of administration. Forexample, in certain embodiments, unless indicated otherwise, thecompound is administered by oral administration, injection, ortransdermal administration. In a preferred embodiment, the compound isadministered orally.

In certain aspects, the therapeutic agents provided herein arecyclically administered to a patient. Cycling therapy involves theadministration of an active agent for a period of time, followed by arest (i.e., discontinuation of the administration) for a period of time,and repeating this sequential administration. Cycling therapy can reducethe development of resistance to one or more of the therapies. Theseregimens can avoid or reduce the side effects of one of the therapies,and/or improve the efficacy of the treatment.

Consequently, in another aspect, a compound provided herein isadministered daily in a single or divided doses in a four to six weekcycle with a rest period of about a week or two weeks. Cycling therapyfurther allows the frequency, number, and length of dosing cycles to beincreased. Thus, another aspect encompasses the administration of acompound provided herein for more cycles than are typical when it isadministered alone. In yet another aspect, a compound provided herein isadministered for a greater number of cycles than would typically causedose-limiting toxicity in a patient to whom a second active ingredientis not also being administered.

In another aspect, a compound provided herein is administered daily andcontinuously for three or four weeks at a dose of from about 0.1 mg toabout 500 mg per day, followed by a rest of one or two weeks. In otherembodiments, the dose can be from about 1 mg to about 500 mg, from about0.1 mg to about 150 mg, from about 1 mg to about 300 mg, from about 10mg to about 100 mg, from about 0.1 mg to about 50 mg, from about 1 mg toabout 50 mg, from about 10 mg to about 50 mg, from about 20 mg to about30 mg, or from about 1 mg to about 20 mg, followed by a rest.

In another aspect, a compound provided herein and a second activeingredient are administered orally or parenterally, with administrationof the compound provided herein occurring prior to (e.g., about 30 to 60minutes) the second active ingredient, during a cycle of four to sixweeks. In certain embodiments, the compound and second active agent areadministered as a single dosage or they are administered separately. Inanother aspect, the combination of a compound provided herein and asecond active ingredient is administered by intravenous infusion overabout 90 minutes every cycle.

Typically, the number of cycles during which the combination treatmentis administered to a patient will be from about one to about 24 cycles,from about two to about 16 cycles, or from about three to about fourcycles.

Combination Therapy

A compound provided herein, or a pharmaceutically acceptable saltthereof, can be combined with other pharmacologically active compounds(“second active agents”) in methods and compositions provided herein.Certain combinations may work synergistically in the treatment ofparticular types of diseases or disorders, and conditions and symptomsassociated with such diseases or disorders. A compound provided herein,or a pharmaceutically acceptable salt thereof, can also work toalleviate adverse effects associated with certain second active agents,and vice versa.

One or more second active ingredients or agents can be used in themethods and compositions provided herein. Second active agents can belarge molecules (e.g., proteins) or small molecules (e.g., syntheticinorganic, organometallic, or organic molecules).

In certain embodiments, the combination therapy comprises adeuterium-enriched compound described herein and a second therapeuticagent for treating a metabolic disorder, such as metformin, a dipeptidylpeptidase IV inhibitor (e.g., sitagliptin, vildagliptin, or the like), astatin (e.g., a HMG-CoA reductase inhibitor, such as atorvastatin,cerivastatin, fluvastatin, lovastatin, mevastatin, simvastatin,rosuvastatin, pravastatin, or combination thereof), a GLP-1 agonist, aGLP-2 agonist, or an SGLT2 inhibitor. As appreciated, the combinationtherapy may comprising more than two therapeutic agents, such as where acombination of a deuterium-enriched compound described herein and atleast two of the aforementioned agents for treating a metabolic disorderare administered to the patient.

In certain other embodiments, the combination therapy comprises adeuterium-enriched compound described herein and a diuretic agent, suchas hydrochlorothiazide.

In certain other embodiments, the combination therapy comprises adeuterium-enriched compound described herein and a second therapeuticagent for treating hypertension, diabetes, or an inflammatory disorder.The second therapeutic agent may be one that limits the activity of therenin-angiotensin system, such as an angiotensin converting enzymeinhibitor (e.g., an ACE inhibitor, such as ramipril, captopril,enalapril, or the like), an angiotensin receptor blocker (e.g.,candesartan, losartan, olmesartan, or the like), or a renin inhibitor.Alternatively, the second therapeutic agent may limit hypertension byalternate means, such as a beta-adrenergic receptor blocker or calciumchannel blocker (e.g., amlodipine).

In certain other embodiments, the combination therapy comprises adeuterium-enriched compound described herein and a glucocorticoidagonist. Such combination therapy may be particularly useful fortreating an inflammatory disorder, such as therapy for suppressing animmune response, preventing transplant rejection, and treatingautoimmune disease. Exemplary disorders include, for example, rheumatoidarthritis, lupus, myasthenia gravis, muscular dystrophy vasculitis,multiple sclerosis, chronic obstructive pulmonary disease (COPD),inflammatory bowel disease, treatment of acute allergic reactions, andtransplant rejection. In certain other embodiments, the combinationtherapy comprises a deuterium-enriched compound described herein and asecond therapeutic agent for treating a kidney disease. Exemplary suchsecond therapeutic agents include those that increase cAMP or comprise abeta-adrenergic agonist. Exemplary beta-adrenergic agonists include, forexample, a beta-1-adrenergic agonist, a beta-2-adrenergic agonist, abeta-3-adrenergic agonist, or a combination thereof. In certainembodiments, the second therapeutic agent is noradrenaline,isoprenaline, dobutamine, salbutamol, levosalbutamol, terbutaline,pirbuterol, procaterol, metaproterenol, fenoterol, bitolterol mesylate,salmeterol, formoterol, bambuterol, clenbuterol, indacaterol, L-796568,amibegron, solabegron, isoproterenol, albuterol, metaproterenol,arbutamine, befunolol, bromoacetylalprenololmenthane, broxaterol,cimaterol, cirazoline, denopamine, dopexamine, epinephrine, etilefrine,hexoprenaline, higenamine, isoetharine, isoxsuprine, mabuterol,methoxyphenamine, nylidrin, oxyfedrine, prenalterol, ractopamine,reproterol, rimiterol, ritodrine, tretoquinol, tulobuterol, xamoterol,zilpaterol, zinterol, or a pharmaceutically acceptable salt thereof; ora combination of any of the foregoing.

In certain other embodiments, the combination therapy comprises adeuterium-enriched compound described herein and a second therapeuticagent for treating cancer. Exemplary second therapeutic agents fortreating cancer include, for example, an alkylating agent, ananti-metabolite (i.e., a molecule that impedes DNA and/or RNAsynthesis), an anti-microtubule agent, a topoisomerase inhibitor, acytotoxic antibiotic, a tyrosine kinase inhibitor, an inhibitor of tumornecrosis factor alpha, anti-neoplastic radiation therapy, or aProgrammed Death protein-1 (PD-1) modulator (e.g., an inhibitor). Incertain embodiments, the second therapeutic agent for treating cancer isazacitidine, azathioprine, bleomycin, carboplatin, capecitabine,carmustine, cisplatin, chlorambucil, cyclophosphamide, cytarabine,dacarbazine, daunorubicin, docetaxel, doxifluridine, doxorubicin,epirubicin, epothilone, etoposide, fluorouracil, fulvestrant,gemcitabine, hydroxyurea, idarubicin, imatinib, lomustine,mechlorethamine, mercaptopurine, methotrexate, mitoxantrone,oxaliplatin, paclitaxel, pemetrexed, procarbazine, raloxifene,teniposide, temozolomide, tamoxifen, toremifene, valrubicin,vinblastine, vincristine, vindesine, vinorelbine, or a pharmaceuticallyacceptable salt thereof.

In yet other embodiments, the second therapeutic agent for treatingcancer is abraxane; acivicin; aclarubicin; acodazole hydrochloride;acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantroneacetate; amrubicin; amsacrine; anastrozole; anthramycin; asparaginase;asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa;bicalutamide; bisantrene hydrochloride; bisnafide dimesylate: bizelesin;bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefmgol: celecoxib;chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; de/.aguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatm;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; herceptin; hydroxyurea;idarubicin hydrochloride; ifosfamide; ilmofosine; iproplatin;irinotecan; irinotecan hydrochloride; lanreotide acetate; lapatinib;letrozole; leuprolide acetate; liarozole hydrochloride; lometrexolsodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine;mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate;melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin;mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride;mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran;paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; portiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; romidepsin; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; a stem cell treatment;streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium;taxotere; tegafur; teloxantrone hydrochloride; temoporfin; teniposide;teroxirone; testolactone; thiamiprine; thioguanine; thiotepa;tiazofurin; tirapazamine; toremifene citrate; trestolone acetate;triciribine phosphate; trimetrexate; trimetrexate glucuronate;triptorelin; tubulozole hydrochloride; uracil mustard; uredepa;vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate;vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; or zorubicinhydrochloride.

Administration of a compound provided herein, or a pharmaceuticallyacceptable salt thereof, and the second active agent(s) to a patient canoccur simultaneously or sequentially by the same or different routes ofadministration. The suitability of a particular route of administrationemployed for a particular active agent will depend on the active agentitself (e.g., whether it can be administered orally without decomposingprior to entering the blood stream) and the disease being treated. Oneroute of administration for compounds provided herein is oral. Routes ofadministration for the second active agents or ingredients are known tothose of ordinary skill in the art. See, e.g., Physicians' DeskReference (60^(th) Ed., 2006).

IV. Pharmaceutical Compositions

The invention provides pharmaceutical compositions comprising adeuterium-enriched compound described herein, such as a compound ofFormula I or II, and a pharmaceutically acceptable carrier. In certainembodiments, the pharmaceutical compositions comprise atherapeutically-effective amount of a deuterium-enriched compounddescribed herein, such as a compound of Formula I or II, formulatedtogether with one or more pharmaceutically acceptable carriers(additives) and/or diluents. As described in detail below, thepharmaceutical compositions of the present invention may be speciallyformulated for administration in solid or liquid form, including thoseadapted for the following: (1) oral administration, for example,drenches (aqueous or non-aqueous solutions or suspensions), tablets(e.g., those targeted for buccal, sublingual, and/or systemicabsorption), boluses, powders, granules, pastes for application to thetongue; (2) parenteral administration by, for example, subcutaneous,intramuscular, intravenous or epidural injection as, for example, asterile solution or suspension, or sustained-release formulation; (3)topical application, for example, as a cream, ointment, or acontrolled-release patch or spray applied to the skin; (4)intravaginally or intrarectally, for example, as a pessary, cream orfoam; (5) sublingually; (6) ocularly; (7) transdermally; or (8) nasally.

Pharmaceutical compositions can be used in the preparation ofindividual, single unit dosage forms. Pharmaceutical compositions anddosage forms provided herein comprise a compound provided herein, or apharmaceutically acceptable salt thereof. Pharmaceutical compositionsand dosage forms can further comprise one or more excipients.Additionally, pharmaceutical compositions and dosage forms providedherein can comprise one or more additional active ingredients. Examplesof optional second, or additional, active ingredients are describedabove.

Single unit dosage forms provided herein are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), topical (e.g., eye drops or other ophthalmicpreparations), transdermal or transcutaneous administration to apatient. Examples of dosage forms include, but are not limited to:tablets; caplets; capsules, such as soft elastic gelatin capsules;cachets; troches; lozenges; dispersions; suppositories; powders;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; eye drops or other ophthalmic preparations suitable fortopical administration; and sterile solids (e.g., crystalline oramorphous solids) that can be reconstituted to provide liquid dosageforms suitable for parenteral administration to a patient.

The composition, shape, and type of dosage forms will typically varydepending on their use. For example, a dosage form used in the acutetreatment of a disease may contain larger amounts of one or more of theactive ingredients it comprises than a dosage form used in the chronictreatment of the same disease. Similarly, a parenteral dosage form maycontain smaller amounts of one or more of the active ingredients itcomprises than an oral dosage form used to treat the same disease. Theseand other ways in which specific dosage forms are used will vary fromone another and will be readily apparent to those skilled in the art.See, e.g., Remington's Pharmaceutical Sciences, 18th ed., MackPublishing, Easton Pa. (1990).

The suitability of a particular excipient may depend on the specificactive ingredients in the dosage form. For example, the decomposition ofsome active ingredients may be accelerated by some excipients such aslactose, or when exposed to water. Active ingredients that compriseprimary or secondary amines are particularly susceptible to suchaccelerated decomposition. Consequently, provided are pharmaceuticalcompositions and dosage forms that contain little, if any, lactose orother mono- or disaccharides. As used herein, the term “lactose-free”means that the amount of lactose present, if any, is insufficient tosubstantially increase the degradation rate of an active ingredient.Lactose-free compositions can comprise excipients that are well known inthe art and are listed, for example, in the U.S. Pharmacopeia (USP)25-NF20 (2002). In general, lactose-free compositions comprise activeingredients, a binder/filler, and a lubricant in pharmaceuticallycompatible and pharmaceutically acceptable amounts. In another aspect,lactose-free dosage forms comprise active ingredients, microcrystallinecellulose, pre-gelatinized starch, and magnesium stearate.

Also provided are anhydrous pharmaceutical compositions and dosage formscomprising active ingredients. Anhydrous pharmaceutical compositions anddosage forms can be prepared using anhydrous or low moisture containingingredients and low moisture or low humidity conditions. Pharmaceuticalcompositions and dosage forms that comprise lactose and at least oneactive ingredient that comprises a primary or secondary amine arepreferably anhydrous if substantial contact with moisture and/orhumidity during manufacturing, packaging, and/or storage is expected. Ananhydrous pharmaceutical composition should be prepared and stored suchthat its anhydrous nature is maintained. Accordingly, anhydrouscompositions are, in another aspect, packaged using materials known toprevent exposure to water such that they can be included in suitableformulary kits. Examples of suitable packaging include, but are notlimited to, hermetically sealed foils, plastics, dose containers (e.g.,vials), blister packs, and strip packs.

Also provided are pharmaceutical compositions and dosage forms thatcomprise one or more compounds that reduce the rate by which an activeingredient will decompose. Such compounds, which are referred to hereinas “stabilizers,” include, but are not limited to, antioxidants such asascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. In another aspect, dosage forms comprise a compoundprovided herein in an amount of from about 0.10 to about 500 mg.Examples of dosages include, but are not limited to, 0.1, 1, 2, 5, 7.5,10, 12.5, 15, 17.5, 20, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450,or 500 mg.

In another aspect, dosage forms comprise the second active ingredient inan amount of 1 to about 1000 mg, from about 5 to about 500 mg, fromabout 10 to about 350 mg, or from about 50 to about 200 mg. Of course,the specific amount of the second active agent will depend on thespecific agent used, the diseases or disorders being treated or managed,and the amount(s) of a compound provided herein, and any optionaladditional active agents concurrently administered to the patient.

Pharmaceutical compositions that are suitable for oral administrationcan be provided as discrete dosage forms, such as, but not limited to,tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g.,flavored syrups). Such dosage forms contain predetermined amounts ofactive ingredients, and may be prepared by methods of pharmacy wellknown to those skilled in the art. See generally, Remington'sPharmaceutical Sciences, 18th ed., Mack Publishing, Easton Pa. (1990).

Oral dosage forms provided herein are prepared by combining the activeingredients in an intimate admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration. For example, excipients suitablefor use in oral liquid or aerosol dosage forms include, but are notlimited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

In another aspect, the invention provides oral dosage forms that aretablets or capsules, in which case solid excipients are employed. Inanother aspect, the tablets can be coated by standard aqueous ornon-aqueous techniques. Such dosage forms can be prepared by any of themethods of pharmacy. In general, pharmaceutical compositions and dosageforms are prepared by uniformly and intimately admixing the activeingredients with liquid carriers, finely divided solid carriers, orboth, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms providedherein include, but are not limited to, binders, fillers, disintegrants,and lubricants. Binders suitable for use in pharmaceutical compositionsand dosage forms include, but are not limited to, corn starch, potatostarch, or other starches, gelatin, natural and synthetic gums such asacacia, sodium alginate, alginic acid, other alginates, powderedtragacanth, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, carboxymethyl cellulose calcium, sodiumcarboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose,pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos.2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms provided herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions is, in anotheraspect, present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Disintegrants may be used in the compositions to provide tablets thatdisintegrate when exposed to an aqueous environment. Tablets thatcontain too much disintegrant may disintegrate in storage, while thosethat contain too little may not disintegrate at a desired rate or underthe desired conditions. Thus, a sufficient amount of disintegrant thatis neither too much nor too little to detrimentally alter the release ofthe active ingredients may be used to form solid oral dosage forms. Theamount of disintegrant used varies based upon the type of formulation,and is readily discernible to those of ordinary skill in the art. Inanother aspect, pharmaceutical compositions comprise from about 0.5 toabout 15 weight percent of disintegrant, or from about 1 to about 5weight percent of disintegrant. Disintegrants that can be used inpharmaceutical compositions and dosage forms include, but are notlimited to, agar-agar, alginic acid, calcium carbonate, microcrystallinecellulose, croscarmellose sodium, crospovidone, polacrilin potassium,sodium starch glycolate, potato or tapioca starch, pre-gelatinizedstarch, other starches, clays, other algins, other celluloses, gums, andmixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms include, but are not limited to, calcium stearate, magnesiumstearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol,polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate,talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil,sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zincstearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof.Additional lubricants include, for example, a Syloid® silica gel(AEROSIL200, manufactured by W. R. Grace Co. of Baltimore, Md.), acoagulated aerosol of synthetic silica (marketed by Degussa Co. ofPlano, Tex.), CAB-O-SIL® (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants may be used in an amount of less than about 1 weight percentof the pharmaceutical compositions or dosage forms into which they areincorporated.

In another aspect, the invention provides a solid oral dosage formcomprising a compound provided herein, anhydrous lactose,microcrystalline cellulose, polyvinylpyrrolidone, stearic acid,colloidal anhydrous silica, and gelatin.

Active ingredients provided herein can also be administered bycontrolled release means or by delivery devices that are well known tothose of ordinary skill in the art. Examples include, but are notlimited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899;3,536,809; 3,598,123; 4,008,719, 5,674,533, 5,059,595, 5,591,767,5,120,548, 5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of whichis incorporated in its entirety herein by reference. Such dosage formscan be used to provide slow or controlled-release of one or more activeingredients using, for example, hydroxypropyl methyl cellulose, otherpolymer matrices, gels, permeable membranes, osmotic systems, multilayercoatings, microparticles, liposomes, microspheres, or a combinationthereof to provide the desired release profile in varying proportions.Suitable controlled-release formulations known to those of ordinaryskill in the art, including those described herein, can be readilyselected for use with the active agents provided herein. In anotheraspect, the invention provides single unit dosage forms suitable fororal administration such as, but not limited to, tablets, capsules, gelcaps, and caplets that are adapted for controlled-release.

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial.Administration of a parenteral dosage form bypasses a patient's naturaldefenses against contaminants, and thus, in these aspects, parenteraldosage forms are sterile or capable of being sterilized prior toadministration to a patient. Examples of parenteral dosage formsinclude, but are not limited to, solutions ready for injection, dryproducts ready to be dissolved or suspended in a pharmaceuticallyacceptable vehicle for injection, suspensions ready for injection, andemulsions. Suitable vehicles that can be used to provide parenteraldosage forms are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms. For example, cyclodextrin and its derivativescan be used to increase the solubility of a compound provided herein.See, e.g., U.S. Pat. No. 5,134,127, which is incorporated in itsentirety herein by reference.

Topical and mucosal dosage forms provided herein include, but are notlimited to, sprays, aerosols, solutions, emulsions, suspensions, eyedrops or other ophthalmic preparations, or other forms known to one ofskill in the art. See, e.g., Remington's Pharmaceutical Sciences, 16thand 18th eds., Mack Publishing, Easton Pa. (1980 & 1990); andIntroduction to Pharmaceutical Dosage Forms, 4th ed. , Lea & Febiger,Philadelphia (1985). Dosage forms suitable for treating mucosal tissueswithin the oral cavity can be formulated as mouthwashes or as oral gels.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide topical and mucosal dosage forms encompassedherein are well known to those skilled in the pharmaceutical arts, anddepend on the particular tissue to which a given pharmaceuticalcomposition or dosage form will be applied. In another aspect,excipients include, but are not limited to, water, acetone, ethanol,ethylene glycol, propylene glycol, butane-1,3-diol, isopropyl myristate,isopropyl palmitate, mineral oil, and mixtures thereof to formsolutions, emulsions or gels, which are nontoxic and pharmaceuticallyacceptable. Moisturizers or humectants can also be added topharmaceutical compositions and dosage forms. Examples of additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa.(1980 & 1990).

The pH of a pharmaceutical composition or dosage form may also beadjusted to improve delivery of one or more active ingredients. Also,the polarity of a solvent carrier, its ionic strength, or tonicity canbe adjusted to improve delivery. Compounds such as stearates can also beadded to pharmaceutical compositions or dosage forms to alter thehydrophilicity or lipophilicity of one or more active ingredients so asto improve delivery. In other aspects, stearates can serve as a lipidvehicle for the formulation, as an emulsifying agent or surfactant, oras a delivery-enhancing or penetration-enhancing agent. In otheraspects, salts of the active ingredients can be used to further adjustthe properties of the resulting composition.

In another aspect, the active ingredients provided herein are notadministered to a patient at the same time or by the same route ofadministration. In another aspect, provided are kits which can simplifythe administration of appropriate amounts of active ingredients.

In another aspect, the invention provides a kit comprising a dosage formof a compound provided herein. Kits can further comprise additionalactive ingredients or a pharmacologically active mutant or derivativethereof, or a combination thereof. Examples of the additional activeingredients include, but are not limited to, those disclosed herein.

In other aspects, the kits can further comprise devices that are used toadminister the active ingredients. Examples of such devices include, butare not limited to, syringes, drip bags, patches, and inhalers.

V. Definitions

To facilitate an understanding of the present invention, a number ofterms and phrases are defined below.

The term “compound” refers to a quantity of molecules that is sufficientto be weighed, tested for its structural identity, and to have ademonstrable use (e.g., a quantity that can be shown to be active in anassay, an in vitro test, or in vivo test, or a quantity that can beadministered to a patient and provide a therapeutic benefit).

Unless indicated otherwise, when a D is specifically recited at aposition or is shown in a formula, this D represents a mixture ofhydrogen and deuterium where the amount of deuterium is about 100%(i.e., the abundance of deuterium ranges from greater than 90% up to100%). In certain embodiments, the abundance of deuterium in D is from95% to 100%, or from 97% to 100%.

The term “patient” refers to organisms to be treated by the methods ofthe present invention. Such organisms preferably include, but are notlimited to, mammals (e.g., murines, simians, equines, bovines, porcines,canines, felines, and the like), and most preferably includes humans

As used herein, the term “effective amount” refers to the amount of acompound sufficient to effect beneficial or desired results. Aneffective amount can be administered in one or more administrations,applications or dosages and is not intended to be limited to aparticular formulation or administration route. As used herein, the term“treating” includes any effect, e.g., lessening, reducing, modulating,ameliorating or eliminating, that results in the improvement of thecondition, disease, disorder, and the like, or ameliorating a symptomthereof.

“Therapeutically effective amount” includes an amount of a compound ofthe invention that is effective when administered alone or incombination to treat the desired condition or disorder. “Therapeuticallyeffective amount” includes an amount of the combination of compoundsclaimed that is effective to treat the desired condition or disorder.The combination of compounds can be additive and is preferably asynergistic combination. Synergy, as described, for example, by Chou andTalalay, Adv. Enzyme Regul. 1984, 22:27-55, occurs when the effect ofthe compounds when administered in combination is greater than theadditive effect of the compounds when administered alone as a singleagent. In general, a synergistic effect is most clearly demonstrated atsub-optimal concentrations of the compounds. Synergy can be in terms oflower incidence of adverse side effects and/or toxicity, increasedefficacy, or some other beneficial effect of the combination comparedwith the individual components.

“Pharmaceutically acceptable salts” refer to derivatives of thedisclosed compounds wherein the parent compound is modified by makingacid or base salts thereof. Examples of pharmaceutically acceptablesalts include, but are not limited to, mineral or organic acid salts ofthe basic residues. The pharmaceutically acceptable salts include theconventional quaternary ammonium salts of the parent compound formed,for example, from non-toxic inorganic or organic acids. These salts canbe prepared in situ in the administration vehicle or the dosage formmanufacturing process, or by separately reacting a purified compound ofthe invention in its free base form with a suitable organic or inorganicacid, and isolating the salt thus formed during subsequent purification.For example, such conventional non-toxic salts include, but are notlimited to, those derived from inorganic and organic acids selected from1,2-ethanedisulfonic, 2-acetoxybenzoic, 2-hydroxyethanesulfonic, acetic,ascorbic, benzenesulfonic, benzoic, bicarbonic, bisulfuric, carbonic,citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric,glucoheptonic, gluconic, glutamic, glycolic, glycollyarsanilic,hexylresorcinic, hydrabamic, hydrobromic, hydrochloric, hydroiodic,hydroxymaleic, hydroxynaphthoic, isethionic, lactic, lactobionic,lauric, lauryl sulfonic, maleic, malic, mandelic, methanesulfonic,napsylic, naphthylic, nitric, oleic, oxalic, palmitic, pamoic,pantothenic, phenylacetic, phosphoric, polygalacturonic, propionic,salicylic, stearic, succinic, sulfamic, sulfanilic, sulfuric, tannic,tartaric, toluenesulfonic, and valeric. (See, for example, Berge et al.(1977) “Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19.). In certainembodiments, the pharmaceutically acceptable salt is a hydrochloric acidsalt.

For therapeutic use, salts of the compounds of the present invention arecontemplated as being pharmaceutically acceptable. However, salts ofacids and bases that are non-pharmaceutically acceptable may also finduse, for example, in the preparation or purification of apharmaceutically acceptable compound.

As used herein, the term “pharmaceutical composition” refers to thecombination of an active agent with a carrier, inert or active, makingthe composition especially suitable for diagnostic or therapeutic use invivo or ex vivo.

As used herein, the term “pharmaceutically acceptable carrier” refers toany of the standard pharmaceutical carriers, such as a phosphatebuffered saline solution, water, emulsions (e.g., such as an oil/wateror a water/oil emulsion), and various types of wetting agents. Thecompositions also can include stabilizers and preservatives. Forexamples of carriers, stabilizers and adjuvants, see e.g., Martin,Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton,Pa. [1975].

Throughout the description, where compositions are described as having,including, or comprising specific components, or where processes andmethods are described as having, including, or comprising specificsteps, it is contemplated that, additionally, there are compositions ofthe present invention that consist essentially of, or consist of, therecited components, and that there are processes and methods accordingto the present invention that consist essentially of, or consist of, therecited processing steps.

As a general matter, if a variable is not accompanied by a definition,then the previous definition of the variable controls.

Finally, the invention may be embodied in other specific forms withoutdeparting from the spirit or essential attributes thereof. Thisinvention encompasses all combinations of aspects and embodiments of theinvention noted herein. It is understood that any and all aspects of theinvention may be taken in conjunction with any other aspects and/orembodiments to describe additional aspects. It is also to be understoodthat each individual element of the aspects is intended to be takenindividually as its own independent aspect. Furthermore, any element ofan aspect is meant to be combined with any and all other elements fromany aspect to describe an additional aspect.

EXAMPLES

The invention now being generally described, will be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

Example 1 Preparation Of Racemic Deuterated Pioglitazone,(Rac-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione)

The hydrochloric acid salt ofrac-5-({p-[2-(5-ethyl-2-pyridyl)ethoxy]phenyl}methyl)-1,3-thiazolidine-2,4-dione(i.e., pioglitazone hydrochloride) (1.5 g, 3.8 mmol) was placed in anoven-dried 250 mL round bottomed flask. Per-deuterated dimethylsulfoxide(d₆-DMSO, 18 mL) and triethylamine (1.596 mL, 11.5 mmol, 3 equiv.) wereadded, followed by per-deuterated methanol (d₄-MeOH, 14 mL). Theresulting suspension was stirred at room temperature while monitoring byLC-MS. After 90 hours, d₆-DMSO (12 mL) and d₄-MeOH (16 mL) were added todissolve the remaining solid. After another 18 hours (total 108 hours),LC-MS analysis showed almost complete deuterium incorporation with%D=98.3% at the chiral center. The mixture was concentrated underreduced pressure, then the concentrate was cooled to 0° C. and dilutedwith cold water (200 mL). The white solid that formed was filtered. Thefiltrate was extracted with ether (2×200 mL), and the organic layerswere combined, dried over sodium sulfate (Na₂SO₄), and filtered. Thewhite solid was combined with the filtrate. The solvent was evaporatedunder reduced pressure and the residue was dried overnight in vacuo togive 1.292 g (3.61 mmol) ofrac-5-({p-[2-(5-ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dioneas a white solid. Overall yield: 1.292 g (3.61 mmol, 95%), %D=98% at thechiral center.

Example 2 Isolation of Enantiopure (R)-Deuterated Pioglitazone and(3)-Deuterated Pioglitazone,((5R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dioneand(5S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione)

rac-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(413 mg, 1.155 mmol) was dissolved in 30 mL of acetonitrile and2-propanol (1:1 v/v). Trifluoroacetic acid (TFA, 225 μL) was added andthe enantiomers (2 mL per run) were separated by supercritical fluidchromatography using a ChiralPak AD-H column (21×250 mm) and a mobilephase of 30% acetonitrile:2-propanol (1:1 v/v) in carbon dioxide (CO₂).Peaks were detected by their UV signal at 254 nm. Fractions containingeach enantiomer were pooled and evaporated. Purity and enantiomericexcess (% ee) were determined by supercritical fluid chromatographyusing an analytical ChiralPak AD-H column (4.6×100 mm) and the samemobile phase. Overall yield was 405.3 mg (1.134 mmol, 98%). The absoluteconfiguration of each enantiomer was determined by measurement of itsspecific rotation in dioxane and then comparison with specific rotationdata already published for the enantiomers of pioglitazone anddeuterated pioglitazone (see International Patent ApplicationPublication Nos. WO 2010/015818 and WO 2010/150014). Physicalcharacterization data for each enantiomer is provided below.

(S)-5-({p-[2(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione (i.e., deuterated (S)-pioglitazone): 214.2 mg(0.599 mmol), 99.6% purity (UV, 220 nm), 99.0% ee: LC/MS: 358.26 (M+1)(>99% deuterium incorporation at the chiral center); ¹H NMR (300 MHz,d₆-DMSO) δ (ppm): 8.34 (s, 1 H), 7.55 (d, 1 H, J=7.8 Hz), 7.25 (d, 1 H,J=7.8 Hz), 7.11 (d, 2 H, J=8.7 Hz), 6.84 (d, 2 H, J=8.7 Hz), 4.29 (t,2H, J=6.6 Hz), 3.28 (d, 1H, J=13.2 Hz), 3.12 (t, 2H, J=6.6 Hz), 3.03 (d,1H, J=14.4 Hz), 2.58 (q, 2H, J=7.7 Hz), 1.16 (t, 3H, J=7.5 Hz); specificrotation [α]_(D)=−72.4° (c 0.5, 19° C., dioxane).

(R)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(i.e., deuterated (R)-pioglitazone): 191.1 mg (0.535 mmol), 100% purity(UV, 220 nm), 100% ee: LC/MS: 358.26 (M+1) (>99% deuterium incorporationat the chiral center); ¹H NMR (300 MHz, d₆-DMSO) δ (ppm): 8.34 (s, 1 H),7.55 (d, 1 H, J=7.8 Hz), 7.25 (d, 1 H, J=7.8 Hz), 7.11 (d, 2 H, J=8.7Hz), 6.84 (d, 2 H, J=8.7 Hz), 4.29 (t, 2 H, J=6.6 Hz), 3.28 (d, 1 H,J=13.2 Hz), 3.12 (t, 2 H, J=6.6 Hz), 3.03 (d, 1 H, J=14.4 Hz), 2.58 (q,2 H, J=7.7 Hz), 1.16 (t, 3 H, J=7.5 Hz); specific rotation [α]_(D)=+94.3(c 0.5, 19° C., dioxane).

Example 3 PPARγAgonist Activity

Agonist activity of deuterated pioglitazone towards the peroxisomeproliferator-activated receptor gamma (PPARγ) was evaluated in twoseparate experiments. Experimental procedures and results from the firstexperiment are provided in Part I below. Experimental procedures andresults from the second experiment are provided in Part II below. Adiscussion of the results from each experiment are provided in Part IIIbelow.

Part I—Analysis of PPARγ Agonist Activity of(S)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(d-S-pio) and(R)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(d-R-pio)

Agonist activity of the enantiomers of5-({p-[2-(5-ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dioneat the peroxisome proliferator-activated receptor gamma (PPARγ) wasevaluated in the thyroid receptor-associated protein complex, 220 kDacomponent (TRAP220) PPARγ coactivator recruitment assay performed atCerep (France). Briefly, a mixture of labeled PPARγ and tagged TRAP220coactivator was pre-incubated at room temperature for 30 minutes in thepresence of a PPARγ-targeted fluorescence acceptor and test compound. ATRAP220-targeted fluorescence donor was then added and the mixture wasincubated for 120 minutes at room temperature. Next, the fluorescencesignal was measured and results expressed as a percent of control (10 μMrosiglitazone). A dose response curve was generated for each enantiomerand the experimental data was analyzed using the log(agonist) vs.response (three parameters) non-linear model in GraphPad Prism 6.0(GraphPad Software, Inc., La Jolla, Calif.), with a fixed Hillslope of1.

(S)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(i.e., deuterated (S)-pioglitazone) was the most potent (EC₅₀=707 nM)and gave the highest maximum coactivator recruitment (106%).(R)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(i.e., deuterated (R)-pioglitazone) was less potent (EC₅₀=4.4 μM) withonly 29% maximum coactivator recruitment when compared to rosiglitazone.

Part II—Analysis of PPARγ Agonist Activity ofrac-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-1,3-thiazolidine-2,4-dione(h-rac-pio);(S)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(d-S-pio); and(R)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(d-R-pio)

PPARγ agonist activity of the following compounds was evaluated in thethyroid receptor-associated protein complex, 220 kDa component (TRAP220)PPARγ coactivator recruitement assay performed at Cerep (France):

-   rac-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-1,3-thiazolidine-2,4-dione-   (S)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione-   (R)-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione.

The experimental procedure involved subjecting a mixture of labeledPPARγ and tagged TRAP220 coactivator to pre-incubation with afluorescence acceptor at room temperature for 30 minutes in the presenceof the test compound. A fluorescence donor was then added, and themixture was incubated for 120 minutes at room temperature. Next, thefluorescence signal was measured and results expressed as a percent ofcontrol (10 μM rosiglitazone). A dose response curve was generated foreach enantiomer and the experimental data was analyzed using thelog(agonist) vs. response (three parameters) non-linear model inGraphPad Prism 6.0 (GraphPad Software, Inc., La Jolla, Calif.), with afixed Hillslope of 1 and maximum of 100%.

Experimental results are depicted in FIG. 1 and EC₅₀ values are providedin Table 5 below. d-S-pio was a more potent PPARγ agonist than h-rac-pioand d-R-pio. In this experiment, d-R-pio did not show any agonistactivity at concentrations up to 100 μM.

TABLE 5 Compound EC₅₀ (μM) d-S-pio 3.47 d-R-pio >100 h-rac-pio 4.63

Part III—Discussion of PPARγ Agonist Activity Results from Parts I andII

Experimental results in Parts I and II illustrate the trend that d-S-piois a much more potent agonist of PPARγ than d-R-pio. Differences in thespecific EC₅₀ values from the experiment in Parts I and II areunderstood to reflect typical differences observed in such cell-basedassays between separate executions of the experiments. Such differencesdo not significantly impact characterization of the relative differencein PPARγ agonist activity for compounds tested under the same executionof the experiment.

Example 4 Human and Mouse Plasma Stability Study of(S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-1,3-Thiazolidine-2,4-Dione;(R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-1,3-Thiazolidine-2,4-Dione;andRac-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione

rac-5-({p-[2-(5-Ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione(d-rac-pio; a 1:1 mixture of (S)- and (R)-enantiomers5-({p-[2-(5-ethyl-2-pyridyl)ethoxy]phenyl}methyl)-(5-²H)-1,3-thiazolidine-2,4-dione)(d-rac-pio),(R)-5-({p-[2-ethyl-2-pyridyl)ethoxy]phenyl}methyl)-1,3-thiazolidine-2,4-dione(h-R-pio), and(S)-5-({p-[2-(5-ethyl-2-pyridyl)ethoxy]phenyl}methyl)-1,3-thiazolidine-2,4-dione(h-S-pio) were dissolved in separate solutions of dimethylsulfoxide(DMSO). The stock solutions were diluted 1:49 v/v in C57BL/6 mouse orhuman plasma to 10, 5, and 5 μM concentrations for d-rac-pio, h-S-pio,and h-R-pio respectively. The plasma samples were incubated at 37° C. induplicate (anticoagulant: K3EDTA). Aliquots (20 μL) were removed at t=0,0.25, 0.5, 1, 2, 4, 8, 16, and 32 h, added to 130 μL acetonitrile, andvortexed. Samples were placed at −80° C. until the study was complete.After thawing, a 1:1 acetonitrile:water solution of internal standard(ISTD, d₄-pioglitazone, 1.6 μM) was added. The vortexed samples werecentrifuged and 50 μL of supernatant was dispensed in a 96-well plate.These were further diluted with 200 pt of 0.1% acetic acid inwater:acetonitrile 15:85 v/v.

The samples were analyzed semi-quantitatively by LC/MS-MS in MRM(multiple reaction monitoring) mode for concentrations of h-S-pio,h-R-pio, d-S-pio, and d-R-pio using a chiral column (ChiralPak IE-3,Chiral Technologies, West Chester, Pa.) for the separation ofenantiomers (isocratic eluent: 01.% acetic acid in water/acetonitrile15/85 v/v at 1 mL/min). All peak areas were normalized to the ISTD andpeak areas for the deuterated enantiomers, d-S-pio and d-R-pio, werecorrected for the isotopic peak of the corresponding protonatedenantiomer, if present (an interference of 12.2% of the response for theprotonated compound was determined experimentally). Corrected data wereanalyzed and plotted using Microsoft Excel 2013 (Microsoft Corp,Redmond, Wash.) and the Excel Solver as well as GraphPad Prism 6.0(GraphPad Software LLC, La Jolla, Calif.) where appropriate.

Scheme 3 illustrates the possible reactions in a solution of deuteratedracemic pioglitazone, where the abbreviations d-S, d-R, h-S, and h-Rrepresent d-S-pio, d-R-pio, h-S-pio, and h-R-pio, respectively. Thedeuterium in both enantiomers, d-S and d-R, can be lost by D/H exchangeto give both protonated enantiomers, h-S and h-R with rate constantsk_(DRR), k_(DRS), k_(DSR), and k_(DSS). At the same time, the protonatedenantiomers h-S and h-R can exchange with enantiomerization rateconstants k_(RS) and k_(SR). All four compounds can also degrade withpotentially different degradation rate constants k_(dSd), k_(dRd),k_(nSd), and k_(hRd).

Human and mouse plasma data were analyzed independently. In addition,since the sum of peak areas for all enantiomeric isotopomers(h-S-pio+h-R-pio+d-S-pio+d-R-pio) appeared independent of incubationtime in plasma from both species, degradation was considered negligible.Therefore, degradation rate constants k_(hSd), k_(hRd), h_(dSd), andk_(dRd) were set to 0. Data in each species was analyzed using astepwise approach. Data for the enantiomerization reaction of h-S-pioand h-R-pio were fitted first and independently of each other. Theaverage rate constants, k_(SR) and k_(RS), obtained from these analyseswere used and fixed in the fitting of the stability data of d-rac-pio.Rate constants k_(DSS), k_(DSR), k_(DRS), and k_(DRR) were obtained fromthis final analysis. Half-lives for the 4 enantiomeric isotopomers werethen calculated as t_(1/2)=1 n(2)/k, where k=k_(DSR)+k_(DRS) ork_(DRR)+k_(DRS) for the deuterated enantiomers (d-S-pio and d-R-pio,respectively) and k=k_(SR) or k_(RS) for the protonated enantiomers(h-S-pio and h-R-pio, respectively).

Data analysis was performed in Microsoft Excel 2013, using the SolverGeneralized Reduced Gradient Nonlinear method with central derivativesto minimize the sum of sums of weighted Δ², square of difference betweenISTD-normalized experimental data and calculated value, divided by theexperimental data (both protonated enantiomers or both protonated anddeuterated enantiomers). Calculated concentrations were obtained throughnumerical approximation of differential equations (1) and (2) for thestability studies of h-S-pio and h-R-pio, and equations (3) to (6) forthe stability study of d-rac-pio by the Euler method (equation (7)). Thestep between calculated time points was minimized in order to minimizethe local error (proportional to the square of the step size) and theglobal error (proportional to the step size).

The observed and fitted data are shown in FIGS. 2A-C for stability inhuman plasma. The observed and fitted data are shown in FIGS. 3A-C forstability in mouse plasma. Fitted parameters are presented in Table 6,which provides rate constants and calculated half-lives (t_(1/2)) forthe in vitro stability of h-S-pio, h-R-pio, and d-rac-pio in human andmouse plasma at 37° C. obtained by fitting experimental data toequations 1 to 6; the DXY stand for the D/H exchange reactions fromd-S-pio (X=S) or d-R-pio (X=R) to h-S-pio (Y=S) or h-R-pio (Y=R); SR andRS represent the enantiomerization reaction h-S-pio to h-R-pio andh-R-pio to h-S-pio, respectively.

$\begin{matrix}{\mspace{79mu}{{\frac{d\left\lbrack {h - S} \right\rbrack}{dt} = {{- {k_{SR}\left\lbrack {h - S} \right\rbrack}} + {k_{RS}\left\lbrack {h - R} \right\rbrack}}}\mspace{79mu}{\frac{d\left\lbrack {h - R} \right\rbrack}{dt} = {{k_{SR}\left\lbrack {h - S} \right\rbrack} - {k_{RS}\left\lbrack {h - R} \right\rbrack}}}{\frac{d\left\lbrack {h - S} \right\rbrack}{dt} = {{- {k_{SR}\left\lbrack {h - S} \right\rbrack}} + {k_{RS}\left\lbrack {h - R} \right\rbrack} + {k_{DSS}\left\lbrack {d - S} \right\rbrack} + {k_{DRS}\left\lbrack {d - R} \right\rbrack}}}{\frac{d\left\lbrack {h - R} \right\rbrack}{dt} = {{k_{SR}\left\lbrack {h - S} \right\rbrack} - {k_{RS}\left\lbrack {h - R} \right\rbrack} + {k_{DSR}\left\lbrack {d - S} \right\rbrack} + {k_{DRR}\left\lbrack {d - R} \right\rbrack}}}\mspace{79mu}{\frac{d\left\lbrack {d - S} \right\rbrack}{dt} = {- {\left( {k_{DSS} + k_{DSR}} \right)\left\lbrack {d - S} \right\rbrack}}}\mspace{79mu}{\frac{d\left\lbrack {d - R} \right\rbrack}{dt} = {- {\left( {k_{DRR} + k_{DRS}} \right)\left\lbrack {d - R} \right\rbrack}}}}} & {{Equations}\mspace{14mu} 1\text{-}6}\end{matrix}$

where [h-S], [h-R], [d-S], [d-R] are the concentrations of h-S-pio,h-R-pio, d-S-pio, and d-R-pio, respectively; k_(SR) and k_(RS) are therate constants for the enantiomerization reactions h-S-pio to h-R-pioand h-R-pio to h-S-pio, respectively; k_(DRR), k_(DRS), k_(DSR), andk_(DSS) are the rate constants for the D/H exchange reactions d-S-pio ord-R-pio to h-S-pio or h-R-pio.

[X]_(t2)=[X]_(t1)+(t ₂ −t ₁)[d[X]]_(t1)   Equation 7

where [X]_(ti) is the concentration of h-S-pio, h-R-pio, d-S-pio ord-R-pio at time ti (wherein i=1 or 2, i.e., ti=t1 or t2), t1 is a timeat which [X] is known, t2 is a time at which [X] is calculated, and[d[X]]_(t1) is the calculated value of the differential equation at timet1.

The equilibrium ratio of enantiomers h-R/h-S was approximately 1:1 inhuman plasma. The equilibrium ratio of enantiomers h-R/h-S wasapproximately 1.25:1 in mouse plasma. The effect of deuteriumincorporation was different for the two enantiomers. For example, anapproximately two-fold increased half-life was observed for d-R-pio vs.h-R-pio. However, the half-life of d-S-pio was approximately the same asthe half-life for h-S-pio.

TABLE 6 Species Compound DSS DSR DRS DRR SR* RS* human d-rac-pio k (h⁻¹)0.209 0.162 0.166 0.0273 0.433 0.440 t_(1/2) (h) 1.9  3.6  1.6   1.6  h-S-pio k (h⁻¹) — — — — 0.427 0.434 t_(1/2) (h) — — 1.6   1.6   h-R-piok (h⁻¹) — — — — 0.439 0.447 t_(1/2) (h) — — 1.6   1.6   mouse d-rac-piok (h⁻¹) 1.038 0.118 0.141 0.218 0.924 0.743 t_(1/2) (h) 0.60 1.93 0.75 0.9   h-S-pio k (h⁻¹) — — — — 0.979 0.792 t_(1/2) (h) — — 0.7   0.9  h-R-pio k (h⁻¹) — — — — 0.867 0.694 t_(1/2) (h) — — 0.8   1.0   *:enantiomerization rate constant used in analysis of stability of d-rac =average of enantiomerization rate constants obtained by fitting data forstability of h-S and h-R

Example 5 Monoamine Oxidase B (MAO-B) Inhibition Study of(S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-S-pio) and(R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-R-pio)

Separate dimethylsulfoxide (DMSO) stock solutions of d-S-pio and d-R-piowere serially diluted in DMSO then mixed with a solution containing 2.5mU of human recombinant monoamine oxidase B in 90 mM4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffercontaining 4.5% glycerol and 9% DMSO. The mixtures were incubated at 22°C. for 5 min. Substrate (D-luciferin derivative) was added, and then themixture was incubated at 37° C. for 60 min. The reaction was stopped byaddition of the detection reagent containing luciferase. Luminescencewas read after standing for 60 min at room temperature.

Experiments were performed in duplicate and a positive control(deprenyl) was used to confirm the validity of the assay. Results wereexpressed as a percentage of the luminescence of the control(enzyme+substrate). IC₅₀ values were obtained by fitting experimentaldata (mean % luminescence as function of concentration) to the Hillequation with variable slope using non-linear regression analysis.

A greater than 5-fold difference in inhibition efficacy was observedbetween d-R-pio and d-S-pio. The results showed d-S-pio to have anIC₅₀=7.6 μM. The results showed d-R-pio to have an IC₅₀=1.4 μM.

Example 6 Effect ofRac-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-1,3-Thiazolidine-2,4-Dione(h-rac-pio);(S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-S-pio); and(R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-R-pio) on Mitochondrial Respiration in C2C12 Cells

The effect of treatment of intact C2C12 cells with h-rac-pio, d-S-pio,and d-R-pio on respiration was evaluated under mitochondrial stressconditions in the presence of sodium pyruvate, in an experimental setupsimilar to that reported by Divakurani et al. in Proc. Natl. Acad. Sci.110 (2013), 5422-5427. Rosiglitazone (10 and 30 μM) and themitochondrial pyruvate carrier inhibitor UK-5099 (300 nM) were used aspositive controls.

On the day before the assay, C2C12 cells were subcultured in XF96microplates at a density of 20,000 cells per well. After overnightincubation (37° C., 5% CO₂), the cells were washed 3 times with assaymedium (Seahorse medium (Seahorse Bioscience, North Billerica, Mass.)containing 10 mM sodium pyruvate, pH 7.4). Compounds, d-S-pio, d-R-pio,or h-rac-pio (3, 10, or 30 μM final concentration), were added to thewells and the plates were incubated at 37° C. (without CO₂). A fullmitochondrial stress test was performed on the XF96 Extracellular FluxAnalyzer (Seahorse Biosciences), including injection of the ATP synthaseinhibitor oligomycin (3.5 μM final), the oxidative phosphorylationuncoupling agent FCCP (carbonyl cyanide4-(trifluoromethoxy)phenylhydrazone, 1 μM final concentration), and thecomplex III inhibitor antimycin A (2.5 μM final). Incubation times of15, 30, or 90 min were used prior to measurement of maximal respiration,i.e., until addition of FCCP. Injection of FCCP always occurred 15 minafter the start of the respirometry assay. Maximal respiration as oxygenconsumption rates (pmoles O₂/min) was measured for each compound, ateach concentration, and each time point in three separate experiments.

Results for the incubation with compounds at 30 μM concentration arepresented in FIG. 4, which shows maximal respiration as oxygenconsumption rate (OCR in pmoles O₂/min) of C2C12 cells treated withh-rac-pio, d-S-pio, or d-R-pio at 30 μM for 15, 30, or 90 minutescompared to the OCR in vehicle-treated cells (average of all repeats andtime points); statistical analysis: one-way ANOVA with Newman-Keulspost-test; *P<0.05; n.s. means not statistically significant. Bothh-rac-pio and d-R-pio inhibited maximal respiration while no significantmeasurable effect was observed for d-S-pio.

Example 7 Pharmacokinetics (PK) ofRac-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-1,3-Thiazolidine-2,4-Dione(h-rac-pio);(R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-R-pio); and(S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-S-pio) Part I—Experimental Procedure

Male C57BL/6 mice (8-10 weeks of age) were randomly divided into 3groups of n=24 animals and administered 30 mg/kg h-rac-pio, 15 mg/kgd-S-pio, or 15 mg/kg d-R-pio (in a 0.25% carboxymethylcellulose solutionprepared daily and used within 1 h of preparation) by oral gavage once aday for 5 days. Blood samples (˜0.5 mL) were collected in K2EDTA tubeson day 5 from n=3 animals per group per time point: pre-dose or 0.25,0.5, 1, 2, 4, 8, or 24 h post-dose by retrobulbar bleeding under lightanesthesia (isoflurane). Animals were then euthanized. Plasma wasseparated by centrifugation and stored at −80° C. until analyzed.

Samples were processed and analyzed by chiral HPLC/MS-MS (ISTD:d₄-pioglitazone) as described in Example 4. Peak areas were normalizedto the peak area of the ISTD and normalized peak areas for deuteratedenantiomers d-S-pio and d-R-pio were corrected for interference from theisotopic peak of the corresponding protonated enantiomer. Concentrationswere calculated by interpolation on standard curves generated from mouseplasma samples spiked with known concentrations of the pure analytes.Data was plotted in Excel 2013 (Microsoft Corp, Redmond, Wash.) andanalyzed within Excel using the PKSolver add-in (version 2.0, asdescribed in Zhang Y. et al. in Comput. Methods Programs Biomed. 99(2010), 306-314) to determine PK parameters including exposure (as areaunder the curve, AUC) and elimination half-life (t_(1/2)).

Part II—Results

PK profiles determined using the above procedure are shown in FIGS.5A-C. Selected PK parameters (C_(max), t_(max), AUC_(0-inf), andt_(1/2)) for the enantiomers of protonated and deuterated pioglitazonein mice after oral gavage of h-rac-pio, d-S-pio, or d-R-pio arepresented in Table 7.

Exposure (as area under the curve, AUC) to the enantiomers of h-rac-piowas stereoselective in animals dosed with h-rac-pio (1:1 mixture ofh-S-pio and h-R-pio) resulting in a 4:1 ratio of h-S-pio to h-R-pio.Dosing d-R-pio resulted in a 10-fold decrease in exposure to the(S)-enantiomer, while exposure to the (R)-enantiomer decreased by only1.7-fold. Dosing d-R-pio resulted in a reversal of the relative exposure(S:R=3:5) compared to what was observed in mice dosed with h-rac-pio.Similarly, dosing d-S-pio resulted in a 7-fold decrease in exposure tothe (R)-enantiomer (vs. dosing h-rac-pio) while exposure to the(S)-enantiomer decreased by 1.5-fold.

TABLE 7 C_(max) (ng/mL) t_(max) (h) Compound h-S h-R d-S d-R h-S h-R d-Sd-R h-rac-pio 20433 7510 — — 1   0.5  — — d-S-pio  2183  574 17133 — 4  0.25 1   — d-R-pio  2127  805   147  6360 4   1    1   1   AUC_(0-inf)(ng.h/mL) t_(1/2) (h) Compound h-S h-R d-S d-R h-S h-R d-S d-R h-rac-pio154240 39702 — — 2.3 2.5 — — d-S-pio  22344  5865 82708 — 3.1 3.2 2.3 —d-R-pio  14036  4850   375 18892 2.5 2.5 0.8 1.8

Example 8 Effect ofRac-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-1,3-Thiazolidine-2,4-Dione(h-rac-pio);(R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-R-pio);(S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-S-pio) on Diabetes and Inflammation in the DB/DB Mouse Model of TypeII Diabetes Part I—Experimental Procedure

Transgenic db/db mice (Jackson Laboratory, Bar Harbor, Me.; n=8 per dosegroup, 8 weeks of age at study start) were dosed daily by oral gavagefor 10 days with vehicle (0.25% carboxymethylcellulose in water),h-rac-pio (30 mg/kg), d-S-pio (20 mg/kg), or d-R-pio (20 mg/kg). Dosingsolutions were prepared daily and used within 1 h of preparation. Bodyweights were measured throughout the study. Non-fasting blood glucose(10 μL by tail snip) was measured 4 hours prior to first dose, andanimals were sorted and assigned to groups based on this measure. Micewere also bled throughout the study through tail snip for blood glucosedetermination at 1 h post-first dose, 1 h post-fifth dose, 1 hpost-eighth dose, and 1 h post-last dose. Animals were euthanized 1 hpost-last dose on day 10. A terminal blood sample was collected fordetermination of serum levels of cholesterol, triglycerides, free fattyacids, adiponectin, serum amyloid A, and cytokines IL-1β, IL-6, TNFα,and MCP-1, using standard analytical methods. Quantitative results wereanalyzed by statistical methods using, as appropriate, one- or two-wayANOVA or Kruskal-Wallis tests for differences followed by Sidak's,Dunnett's, or Dunn's post-test to determine differences vs.vehicle-treated control animals. All graphs were generated with GraphPadPrism version 6.0.2 (GraphPad Software, Inc., La Jolla, Calif.).

Part II—Results

No significant differences in body weight were observed between dosegroups. No overt signs of toxicity were seen in any animal of any dosegroup.

Blood glucose data are presented in FIG. 6, which shows the amount ofnon-fasted blood glucose (ng/mL) in db/db mice dosed daily by oralgavage with vehicle (hollow circles), 20 mg/kg d-S-pio (filledtriangles), 20 mg/kg d-R-pio (filled squares), or 30 mg/kg h-rac-pio(hollow triangles) measured on day one at 4 hours pre-first dose, and ondays one, eight, and ten, 1 hour post-dose (2-way repeated measuresANOVA with Sidak post-test, ***P<0.001).

Effects of h-rac-pio, d-S-pio, and d-R-pio on metabolic diseasebiomarkers, adiponectin, and inflammatory biomarkers are summarized inFIGS. 7A-C, which illustrate the effect of 10-day, daily oraladministration of vehicle, d-S-pio (20 mg/kg), d-R-pio (20 mg/kg), orh-rac-pio (30 mg/kg) to db/db mice on certain metabolic diseasebiomarkers (i.e., insulin, cholesterol, triglycerides, non-essentialfatty acids), adiponectin, and inflammatory biomarkers (i.e., IL-1β,IL-6, TNF-α, MCP-1, and serum amyloid A) (Kruskal-Wallis test withDunn's post-test against vehicle, *P<0.05, **P<0.01, ***P<0.001, and****P<0.0001, for insulin, cholesterol, triglycerides, non-essentialfatty acids, adiponectin, MCP-1; one-tailed unpaired t-test againstvehicle, *P<0.05, for IL-1β, IL-6, TNF-α, and serum amyloid A).

The results show that both d-S-pio and d-R-pio decreased non-fastedblood glucose to approximately the same extent as h-rac-pio. The samewas observed for serum triglycerides, while a similar trend was observedfor non-essential fatty acids.

Only d-S-pio and h-rac-pio significantly increased adiponectin. Onlyh-rac-pio significantly decreased insulin. Inflammatory biomarkers weresimilarly decreased by d-R-pio and h-rac-pio: trending for cytokinesIL-1β, IL-6, and TNF-α, and statistically significant for serum amyloidA.

Example 9 Effect ofRac-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-1,3-Thiazolidine-2,4-Dione(h-rac-pio);(R)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-R-pio); and(S)-5-({p-[2-(5-Ethyl-2-Pyridyl)Ethoxy]Phenyl}Methyl)-(5-²H)-1,3-Thiazolidine-2,4-Dione(d-S-pio) on Body Weight Gain in Male C57BL/6J Mice on a Standard DietPart I—Experimental Procedure

Male C57BL/6J mice (about 28 g body weight) were acclimatedsingly-housed for one week to a reverse light/dark cycle with freeaccess to water and standard maintenance diet. Animals were then handleddaily for one week followed by another 7 days of twice daily dosing withvehicle (0.5% methylcellulose in water) by oral gavage. After thehabituation period the mice were allocated to 1 of 4 dosing groups (n=12to 14 animals per group; 14 for vehicle control) on the basis of bodyweight, and food and water intake. Animals were then dosed twice dailyby oral gavage with vehicle, h-rac-pio (30 mg/kg overall daily dose,i.e., two doses of 15 mg/kg daily), d-S-pio (15 mg/kg overall dailydose, i.e., two doses of 7.5 mg/kg daily), or d-R-pio (15 mg/kg overalldaily dose, i.e., two doses of 7.5 mg/kg daily) for 11 days at thebeginning and end of the dark phase. Dosing solutions (0.5%methylcellulose in water adjusted to pH˜7 for d-S-pio and d-R-pio) wereprepared twice daily and used within 1 h of preparation. Body weightswere measured twice daily, prior to each dosing, to adjust individualdosing volumes to actual body weights. The morning body weights wererecorded and are presented in FIG. 8 as percent body weight differenceversus day 1.

Part II—Results

Dosing with h-rac-pio and d-S-pio resulted in a statisticallysignificant increase in percent body weight difference compared todosing with vehicle. Oral gavage with d-R-pio did not increase percentbody weight difference over vehicle. Results are depicted graphically inFIG. 8, which shows the effect of twice daily dosing with vehicle(hollow circles), h-rac-pio (30 mg/kg overall daily dose, hollowtriangles), d-S-pio (15 mg/kg overall daily dose, filled triangles), andd-R-pio (15 mg/kg overall daily dose, filled squares) on body weight,expressed as percent body weight difference versus day 1 body weight(mean±SEM) in male C57BL/6J mice (n=14 mice in vehicle group, n=12 eachin h-rac-pio, d-S-pio, and d-R-pio groups; ANOVA statistical analysiswith multiple comparison Dunnett's post-test, *P<0.05, **P<0.01,***P<0.001).

INCORPORATION BY REFERENCE

All references listed herein are individually incorporated in theirentirety by reference.

Equivalents

Numerous modifications and variations of the invention are possible inlight of the above teachings. It is therefore to be understood thatwithin the scope of the appended claims, the invention may be practicedotherwise that as specifically described herein.

1-87. (canceled)
 88. A compound represented by

or a pharmaceutically acceptable salt thereof, having an optical purityof at least 75% enantiomeric excess, wherein Z is H or D, provided thatthe abundance of deuterium in Z is at least 30%.
 89. The compound ofclaim 88, wherein the abundance of deuterium is at least 75%.
 90. Thecompound of claim 88, wherein the abundance of deuterium is at least90%.
 91. The compound of claim 88, having an optical purity of at least90% enantiomeric excess.
 92. A compound represented by

or a pharmaceutically acceptable salt thereof, having an optical purityof at least 75% enantiomeric excess, wherein Z is H or D, provided thatthe abundance of deuterium in Z is at least 30%.
 93. The compound ofclaim 92 wherein the abundance of deuterium is at least 75%.
 94. Thecompound of claim 92, wherein the abundance of deuterium is at least90%.
 95. The compound of claim 92, having an optical purity of at least90% enantiomeric excess.
 96. A method of treating a neurologicaldisorder selected from the group consisting of Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis. Friedreich's ataxia,autism spectrum disorder, depression, mild cognitive impairment, Downsyndrome, neurodegeneration, adrenoleukodystrophy, Huntington's disease,stroke, traumatic brain injury, substance abuse, spinal cord injury,neuronal injury, major depression or bipolar disorder comorbid withmetabolic syndrome, and a neurological disorder caused by functionalmitochondrial impairment, comprising administering to a patient in needthereof a therapeutically effective amount of the compound of claim 88.97. A method of treating a disorder selected from the group consistingof cancer, a metabolic disorder, a symptom of hepatitis, acardiovascular disease, polycystic ovary syndrome, and a skin defectcaused by exposure to ultraviolet radiation, comprising administering toa patient in need thereof a therapeutically effective amount of thecompound of claim
 88. 98. The method of claim 97, wherein the metabolicdisorder is non-alcoholic fatty liver disease, viral hepatitis, livercirrhosis, liver fibrosis, diabetic retinopathy, diabetic neuropathy,diabetic nephropathy, beta cell depletion, insulin resistance in apatient with congenital adrenal hyperplasia treated with aglucocorticoid, dysmetabolism in peritoneal dialysis patients, reducedinsulin secretion, improper distribution of brown fat cells and whitefat cells, obesity, or improper modulation of leptin levels.
 99. Amethod of treating a neurological disorder selected from the groupconsisting of Alzheimer's disease, Parkinson's disease, amyotrophiclateral sclerosis, Friedreich's ataxia, autism spectrum disorder,depression, mild cognitive impairment, Down syndrome, neurodegeneration,adrenoleukodystrophy, Huntington's disease, stroke, traumatic braininjury, substance abuse, spinal cord injury, neuronal injury, majordepression or bipolar disorder comorbid with metabolic syndrome. and aneurological disorder caused by functional mitochondrial impairment,comprising administering to a patient in need thereof a therapeuticallyeffective amount of the compound of claim
 92. 100. A method of treatinga disorder selected from the group consisting of cancer, a metabolicdisorder, a symptom of hepatitis, a cardiovascular disease, polycysticovary syndrome, and a skin defect caused by exposure to ultravioletradiation, comprising administering to a patient in need thereof atherapeutically effective amount of the compound of claim
 92. 101. Themethod of claim 100, wherein the metabolic disorder is non-alcoholicfatty liver disease, viral hepatitis, liver cirrhosis, liver fibrosis,diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, betacell depletion, insulin resistance in a patient with congenital adrenalhyperplasia treated with a glucocorticoid, dysmetabolism in peritonealdialysis patients, reduced insulin secretion, improper distribution ofbrown fat cells and white fat cells, obesity, or improper modulation ofleptin levels.
 102. A method of synthesizing the compound F1, the methodcomprising reaction of E1 with d₆-DMSO and triethylamine followed byd₄-MeOH